Abuse resistant lysine amphetamine compounds

ABSTRACT

The present invention describes compounds, compositions and methods of using the same comprising lysine covalently attached to amphetamine. These compounds and compositions are useful for reducing or preventing abuse and overdose of amphetamine. These compounds and compositions find particular use in providing an abuse-resistant alternative treatment for certain disorders, such as attention deficit hyperactivity disorder (ADHD), ADD, narcolepsy, and obesity. Oral bioavailability of amphetamine is maintained at therapeutically useful doses. At higher doses bioavailability is substantially reduced, thereby providing a method of reducing oral abuse liability. Further, compounds and compositions of the invention decrease the bioavailability of amphetamine by parenteral routes, such as intravenous or intranasal administration, further limiting their abuse liability.

CROSS REFERENCE RELATED APPLICATIONS

This application claims benefit under 35 U.S.C. 119(e) to U.S.provisional application No. 60/473,929 filed May 29, 2003 andprovisional application No. 60/567,801 filed May 5, 2004, both of whichare hereby incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

(i) Field of the Invention

The invention relates to amphetamine compounds, compositions and methodsof delivery and use comprising amphetamine covalently bound to achemical moiety.

The invention relates to compounds comprised of amphetamine covalentlybound to a chemical moiety in a manner that diminishes or eliminatespharmacological activity of amphetamine until released. The conjugatesare stable in tests that simulate procedures likely to be used byillicit chemists in attempts to release amphetamine. The inventionfurther provides for methods of therapeutic delivery of amphetaminecompositions by oral administration. Additionally, release ofamphetamine following oral administration occurs gradually over anextended period of time thereby eliminating spiking of drug levels. Whentaken at doses above the intended prescription, the bioavailability ofamphetamine, including peak levels and total amount of drug absorbed, issubstantially decreased. This decreases the potential for amphetamineabuse which often entails the use of extreme doses (1 g or more a day).The compositions are also resistant to abuse by parenteral routes ofadministration, such as intravenous “shooting”, intranasal “snorting”,or inhalation “smoking”, that are often employed in illicit use. Theinvention thus provides a stimulant based treatment for certaindisorders, such as attention deficit hyperactivity disorder (ADHD),which is commonly treated with amphetamine. Treatment of ADHD withcompositions of the invention results in substantially decreased abuseliability as compared to existing stimulant treatments.

(ii) Background of the Invention

The invention is directed to amphetamine conjugate compounds,compositions, and methods of manufacture and use thereof. In particular,the invention is directed to an anti-abuse/sustained release formulationwhich maintains its therapeutic effectiveness when administered orally.The invention further relates to formulations which diminish or reducethe euphoric effect while maintaining therapeutically effective bloodconcentrations following oral administration.

Amphetamine is prescribed for the treatment of various disorders,including attention deficit hyperactivity disorder (ADHD), obesity andnarcolepsy. Amphetamine and methamphetamine stimulate the centralnervous system and have been used medicinally to treat ADHD, narcolepsyand obesity. Because of its stimulating effects amphetamine and itsderivatives (e.g., amphetamine analogues) are often abused. Similarly,p-methoxyamphetamine, methylenedioxyamphetamine,2,5-dimethoxy-4-methylamphetamine, 2,4,5-trimethoxyamphetamine and3,4-methylenedioxymethamphetamine are also often abused.

In children with attention deficit hyperactivity disorder (ADHD), potentCNS stimulants have been used for several decades as a drug treatmentgiven either alone or as an adjunct to behavioral therapy. Whilemethylphenidate (Ritalin) has been the most frequently prescribedstimulant, the prototype of the class, amphetamine (alpha-methylphenethylamine) has been used all along and increasingly so in recentyears. (Bradley C, Bowen M, “Amphetamine (Benzedrine) therapy ofchildren's behavior disorders.” American Journal of Orthopsychiatry 11:92) (1941).

The potential for abuse of amphetamines is a major drawback to its use.The high abuse potential has earned it Schedule II status according tothe Controlled Substances Act (CSA). Schedule II classification isreserved for those drugs that have accepted medical use but have thehighest potential for abuse. The abuse potential of amphetamine has beenknown for many years and the FDA requires the following black boxwarning in the package inserts of products:

AMPHETAMINES  HAVE  A  HIGH  POTENTIAL  FOR ABUSE.   ADMINISTRATION OFAMPHETAMINES FOR PROLONGED PERIODS OF TIME MAY LEAD TO DRUG DEPENDENCEAND MUST BE AVOIDED. PARTICULAR ATTENTION SHOULD BE PAID TO THEPOSSIBILITY OF SUBJECTS OBTAINING AMPHETAMINES FOR NONTHERAPEUTIC USE ORDISTRIBUTION TO OTHERS, AND THE DRUGS SHOULD BE PRESCRIBED OR DISPENSEDSPARINGLY.

Furthermore, recent developments in the abuse of prescription drugproducts increasingly raise concerns about the abuse of amphetamineprescribed for ADHD. Similar to OxyContin, a sustained releaseformulation of a potent narcotic analgesic, Adderall XR® represents aproduct with increased abuse liability relative to the single dosetablets. The source of this relates to the higher concentration ofamphetamine in each tablet and the potential for release of the fullamount of active pharmaceutical ingredient upon crushing. Therefore,like OxyContin, it may be possible for substance abusers to obtain ahigh dose of the pharmaceutical with rapid onset by snorting the powderor dissolving it in water and injecting it. (Cone, E. J., R. V. Fant, etal., “Oxycodone involvement in drug abuse deaths: a DAWN-basedclassification scheme applied to an oxycodone postmortem databasecontaining over 1000 cases.” J Anal Toxicol 27(2): 57-67; discussion 67)(2003).

It has been noted recently that “53 percent of children not takingmedication for ADHD knew of students with the disorder either givingaway or selling their medications. And 34 percent of those being treatedfor the disorder acknowledged they had been approached to sell or tradethem.” (Dartmouth-Hitchcock, 2003) “Understanding ADHD Stimulant Abuse.”http://12.42.224.168/healthyliving/familyhome/jan03familyhomestimulantabuse.htm).In addition, it was reported that students at one prep school obtainedDexedrine and Adderall to either swallow tablets whole or crush andsniff them. (Dartmouth-Hitchcock (2003).

According to the drug enforcement administration (DEA, 2003):

-   -   Methylphenidate and amphetamine can be abused orally or the        tablets can be crushed and snorted or dissolved in water and        injected. The pattern of abuse is characterized by escalation in        dose, frequent episodes of binge use followed by severe        depression and an overpowering desire to continue the use of        these drugs despite serious adverse medical and social        consequences.    -   Rendering this potent stimulant resistant to abuse, particularly        by parenteral routes such as snorting or injecting, would        provide considerable value to this otherwise effective and        beneficial prescription medication.    -   (DEA (2003). “Stimulant Abuse By School Age Children: A Guide        for School Officials.        “http://www.deadiversion.usdoj.gov/pubs/brochures/stimulant/stimulant_abuse.htm).

Typically, sustained release formulations contain drug particles mixedwith or covered by a polymer material, or blend of materials, which areresistant to degradation or disintegration in the stomach and/or in theintestine for a selected period of time. Release of the drug may occurby leeching, erosion, rupture, diffusion or similar actions dependingupon the nature of the polymer material or polymer blend used.Additionally, these formulations are subject to breakdown followingrelatively simple protocols which allows for abuse of the activeingredient.

Conventionally, pharmaceutical manufacturers have used hydrophilichydrocolloid gelling polymers such as hydroxypropyl methylcellulose,hydroxypropyl cellulose or Pullulan to formulate sustained releasetablets or capsules. These polymers first form a gel when exposed to anaqueous environment of low pH thereby slowly diffusing the activemedicament which is contained within the polymer matrix. When the gelenters a higher pH environment such as that found in the intestines,however, it dissolves resulting in a less controlled drug release. Toprovide better sustained release properties in higher pH environments,some pharmaceutical manufacturers use polymers which dissolve only athigher pHs, such as acrylic resins, acrylic latex dispersions, celluloseacetate phthalate, and hydroxypropyl methylcellulose phthalate, eitheralone or in combination with hydrophilic polymers.

These formulations are prepared by combining the medicament with afinely divided powder of the hydrophilic polymer, or the hydrophilic andwater-insoluble polymers. These ingredients are mixed and granulatedwith water or an organic solvent and the granulation is dried. The drygranulation is then usually further blended with various pharmaceuticaladditives and compressed into tablets.

Although these types of formulations have been successfully used tomanufacture dosage forms which demonstrate sustained release properties,these formulations are subject to several shortcomings including unevenrelease and are subject to abuse.

The need exists for an abuse resistant dosage form of amphetamine whichis therapeutically effective. Further the need exists for an amphetaminedosage form which provides sustained release and sustained therapeuticeffect.

SUMMARY OF INVENTION

The invention provides covalent attachment of amphetamine andderivatives or analogs thereof to a variety of chemical moieties. Thechemical moieties may include any substance which results in a prodrugform, i.e., a molecule which is converted into its active form in thebody by normal metabolic processes. The chemical moieties may be forinstance, amino acids, peptides, glycopeptides, carbohydrates,nucleosides, or vitamins.

The chemical moiety is covalently attached either directly or indirectlythrough a linker to the amphetamine. The site of attachment is typicallydetermined by the functional group(s) available on the amphetamine.

In one embodiment of the invention, the chemical moiety is a carrierpeptide as defined herein. The carrier peptide may be attached toamphetamine through the carrier's N-terminus, C-terminus or side chainof an amino acid which may be either a single amino acid or part of alonger chain sequence (i.e. a dipeptide, tripeptide, an oligopeptide ora polypeptide). Preferably, the carrier peptide is (i) an amino acid,(ii) a dipeptide, (iii) a tripeptide, (iv) an oligopeptide, or (v)polypeptide. The carrier peptide may also be (i) a homopolymer of anaturally occurring amino acid, (ii) a heteropolymer of two or morenaturally occurring amino acids, (iii) a homopolymer of a syntheticamino acid, (iv) a heteropolymer of two or more synthetic amino acids,or (v) a heteropolymer of one or more naturally occurring amino acidsand one or more synthetic amino acids. A further embodiment of thecarrier and/or conjugate is that the unattached portion of thecarrier/conjugate may be in a free and unprotected state. Preferably,synthetic amino acids with alkyl side chains are selected from alkyls ofC₁-C₁₇ in length and more preferably from C₁-C₆ in length.

Covalent attachment of a chemical moiety to amphetamine can decrease itspharmacological activity when administered through injection orintranasally. Compositions of the invention, however, provideamphetamine covalently attached to a chemical moiety which remainsorally bioavailable. The bioavailability is a result of the hydrolysisof the covalent linkage following oral administration. Hydrolysis istime-dependent, thereby allowing amphetamine to become available in itsactive form over an extended period of time. In one embodiment, thecomposition provides oral bioavailability which resembles thepharmacokinetics observed for extended release formulations. In anotherembodiment, release of amphetamine is diminished or eliminated whendelivered by parenteral routes.

In one embodiment, the compositions maintain their effectiveness andabuse resistance following the crushing of the tablet, capsule or otheroral dosage form. In contrast, conventional extended releaseformulations used to control the release of amphetamine throughincorporation into matrices are subject to release of up to the entireamphetamine content immediately following crushing. When the content ofthe crushed tablet is injected or snorted, the large dose of amphetamineproduces the “rush” effect sought by addicts.

In one embodiment, the amphetamine is attached to a single amino acidwhich is either naturally occurring or a synthetic amino acid. Inanother embodiment, the amphetamine is attached to a dipeptide ortripeptide, which could be any combination of the naturally occurringamino acids and synthetic amino acids. In another embodiment, the aminoacids are selected from L-amino acids for digestion by proteases.

In another embodiment, the side chain attachment of amphetamine to thepolypeptide or amino acid are selected from homopolymers orheteropolymers of glutamic acid, aspartic acid, serine, lysine,cysteine, threonine, asparagine, arginine, tyrosine, and glutamine.Examples of peptides include, Lys, Ser, Phe, Gly-Gly-Gly, Leu-Ser,Leu-Glu, homopolymers of Glu and Leu, and heteropolymers of(Glu)n-Leu-Ser. In a preferred embodiment, the composition is selectedfrom Lys-Amp, Ser-Amp, Phe-Amp, and Gly-Gly-Gly-Amp.

In another embodiment, the invention provides a carrier and amphetaminewhich are bound to each other but otherwise unmodified in structure.This embodiment may further be described as the carrier having a freecarboxy and/or amine terminal and/or side chain groups other than at thelocation of attachment for the amphetamine. In a preferred embodiment,the carrier, whether a single amino acid, dipeptide, tripeptide,oligopeptide or polypeptide, comprises only naturally occurring aminoacids.

Another embodiment of the invention provides a method for deliveringamphetamine dosage which prevents euphoria, comprising administering toa patient in need a composition formulated for oral dosage comprisingamphetamine covalently attached to a chemical moiety wherein said bloodlevels of amphetamine maintain a therapeutically effect level but do notresult in a euphoric effect.

In another embodiment, the covalent attachment of a chemical moietysubstantially decreases the potential for overdose by decreasing thetoxicity of amphetamine at doses above those considered therapeutic,while maintaining its pharmaceutical activity within a normal doserange. Covalent attachment of the chemical moiety may decrease oreliminate the pharmacological activity of amphetamine. Therefore,restoring activity requires release of the amphetamine from the chemicalmoiety. At higher doses partial or complete saturation of processesresponsible for amphetamine release may be reached thus diminishing oreliminating the release of harmful levels of active amphetamine. Forexample, aspects of pharmacological activity, release, saturation arefurther depicted in FIGS. 1-55.

In another embodiment of the invention, the covalent attachment of achemical moiety substantially decreases the potential for overdose bydecreasing the rate or overall amount of absorption of the amphetaminewhen given at doses above those considered therapeutic.

In another embodiment of the invention, the covalent attachment of achemical moiety substantially decreases the potential for overdose byincreasing the rate or overall amount of clearance of amphetamine whengiven at doses above those considered therapeutic.

Another embodiment provides a method of treating a patient sufferingfrom attention deficit hyperactivity disorder, narcolepsy or obesitycomprising providing, administering, prescribing, etc. compositions ofthe invention.

Another embodiment of the invention provides a method for deliveringamphetamine, comprising providing a patient with a therapeuticallyeffective amount of amphetamine covalently attached to a chemical moietywhich provides a therapeutically bioequivalent AUC when compared toamphetamine alone but does not provide a C_(max) which results ineuphoria when taken orally.

Other objects, advantages and embodiments of the invention are describedbelow and will be obvious from this description and practice of theinvention.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1. Synthesis of amino acid amphetamine conjugates.

FIG. 2. Synthesis of lysine amphetamine conjugate.

FIG. 3. Synthesis of serine amphetamine conjugate.

FIG. 4. Synthesis of phenylalanine amphetamine conjugate.

FIG. 5. Synthesis of triglycine amphetamine conjugate.

FIG. 6. Plasma concentrations of d-amphetamine from individual animalsorally administered d-amphetamine or L-lysine-d-amphetamine.

FIG. 7. Plasma concentrations of d-amphetamine following oraladministration of d-amphetamine sulfate or L-lysine-d-amphetamine (1.5mg/kg d-amphetamine base) to rats (ELISA analysis).

FIG. 8. Plasma concentrations of d-amphetamine following oraladministration of d-amphetamine sulfate or L-lysine-d-amphetamine (3mg/kg d-amphetamine base) to rats (ELISA analysis).

FIG. 9. Plasma concentrations of d-amphetamine following oraladministration of d-amphetamine sulfate or L-lysine-d-amphetamine (6mg/kg d-amphetamine base) to rats (ELISA analysis).

FIG. 10. Plasma concentrations of d-amphetamine at 30-minutes post-dosefor escalating doses of L-lysine-d-amphetamine or d-amphetamine sulfate(ELISA analysis).

FIG. 11. Plasma concentrations of d-amphetamine following oraladministration of L-lysine-d-amphetamine or d-amphetamine sulfate (60mg/kg d-amphetamine base) to rats (ELISA analysis).

FIG. 12. Plasma concentrations of d-amphetamine following intranasaladministration of L-lysine-d-amphetamine or d-amphetamine sulfate (3mg/kg d-amphetamine base) to rats (ELISA analysis).

FIG. 13. Plasma concentrations of d-amphetamine following bolusintravenous administration of L-lysine-d-amphetamine or d-amphetaminesulfate (1.5 mg/kg d-amphetamine base) to rats (ELISA analysis).

FIG. 14. Plasma concentrations of d-amphetamine levels following oraladministration of Dexadrine Spansule capsules, crushed DexadrineSpansule capsules, or L-lysine-d-amphetamine (3 mg/kg d-amphetaminebase) to rats (ELISA analysis).

FIGS. 15A-B. Plasma concentrations of d-amphetamine in ng/mL (FIG. 15A),and in uM (FIG. 15B), following oral administration ofL-lysine-d-amphetamine or d-amphetamine sulfate (1.5 mg/kg d-amphetaminebase) to rats (LC/MS/MS analysis).

FIGS. 16A-B. Plasma concentrations of d-amphetamine in ng/mL (FIG. 16A),and in uM (FIG. 16B), following oral administration ofL-lysine-d-amphetamine or d-amphetamine sulfate (3 mg/kg d-amphetaminebase) to rats (LC/MS/MS analysis).

FIGS. 17A-B. Plasma concentrations of d-amphetamine in ng/mL (FIG. 17A),and in uM (FIG. 17B), following oral administration ofL-lysine-d-amphetamine or d-amphetamine sulfate (6 mg/kg d-amphetaminebase) to rats (LC/MS/MS analysis).

FIGS. 18A-B. Plasma concentrations of d-amphetamine in ng/mL (FIG. 18A),and in uM (FIG. 18B), following oral administration ofL-lysine-d-amphetamine or d-amphetamine sulfate (12 mg/kg d-amphetaminebase) to rats (LC/MS/MS analysis).

FIGS. 19A-B. Plasma concentrations of d-amphetamine in ng/mL (FIG. 19A),and in uM (FIG. 19B), following oral administration of or d-amphetaminesulfate (60 mg/kg d-amphetamine base) to rats (LC/MS/MS analysis).

FIG. 20. Comparative bioavailability (C_(max)) of L-lysine-d-amphetamineand d-amphetamine in proportion to escalating human equivalent doses inrats (mg/kg d-amphetamine base).

FIG. 21. Comparative bioavailability (AUC_(inf)) ofL-lysine-d-amphetamine and d-amphetamine in proportion to escalatingdoses in rats (mg/kg d-amphetamine base).

FIG. 22. Comparative Bioavailability (AUC_(inf)) ofL-lysine-d-amphetamine and d-amphetamine in proportion to escalatinghuman equivalent doses in rats (mg/kg d-amphetamine base).

FIG. 23. Plasma concentrations of d-amphetamine following intranasaladministration of L-lysine-d-amphetamine or d-amphetamine sulfate (3mg/kg d-amphetamine base) to rats (LC/MS/MS analysis).

FIG. 24. Plasma concentrations of d-amphetamine andL-lysine-d-amphetamine in ng/mL (FIG. 24A), and in μM (FIG. 24B),following intranasal administration of L-lysine-d-amphetamine ord-amphetamine sulfate (3 mg/kg d-amphetamine base) to rats (LC/MS/MSanalysis).

FIG. 25. Plasma concentrations of d-amphetamine following bolusintravenous administration of L-lysine-d-amphetamine or d-amphetaminesulfate (1.5 mg/kg d-amphetamine base) to rats (LC/MS/MS analysis).

FIGS. 26A-B. Plasma concentrations of d-amphetamine in ng/mL (FIG. 26A),and in μM (FIG. 26B), following intranasal administration ofL-lysine-d-amphetamine or d-amphetamine sulfate (3 mg/kg d-amphetaminebase) to rats (LC/MS/MS analysis).

FIG. 27. Mean plasma concentration time profile ofL-lysine-d-amphetamine following 30-min intravenous infusion (2 mg/kg)or oral administration of L-lysine-d-amphetamine (2 mg/kg) in consciousmale beagle dogs (n=3).

FIG. 28. Plasma concentration time profile of d-amphetamine following30-min intravenous infusion or oral administration ofL-lysine-d-amphetamine (2 mg/kg) in conscious male beagle dogs (n=3).

FIGS. 29A-B. Mean plasma concentration time profile ofL-lysine-d-amphetamine and d-amphetamine levels in ng/ml (FIG. 29A), andin uM (FIG. 29B), following 30-min intravenous infusion (2 mg/kg) inconscious male beagle dogs (n=3).

FIGS. 30A-B. Mean plasma concentration time profile ofL-lysine-d-amphetamine and d-amphetamine levels in ng/ml (FIG. 30A), andin nM (FIG. 30B), following oral administration ofL-lysine-d-amphetamine (2 mg/kg) in conscious male beagle dogs (n=3).

FIGS. 31A-B. Individual plasma concentration time profile ofL-lysine-d-amphetamine following intravenous administration (FIG. 31A)or oral administration (FIG. 31B) of L-lysine-d-amphetamine in consciousmale beagle dogs. The oral formulation used comprises solution and 0.2mg/mL in water.

FIGS. 32A-B. Individual plasma concentration time profile ofd-amphetamine following intravenous administration (FIG. 32A) or oraladministration (FIG. 32B) of L-lysine-d-amphetamine in conscious malebeagle dogs.

FIG. 33. Plasma concentrations of d-amphetamine following oraladministration of L-lysine-d-amphetamine or d-amphetamine sulfate (1.8mg/kg d-amphetamine base) to male dogs.

FIG. 34. Plasma concentrations of d-amphetamine following oraladministration of L-lysine-d-amphetamine or d-amphetamine sulfate (1.8mg/kg d-amphetamine base) to female dogs.

FIG. 35. Mean blood pressure following intravenous bolus injection ofincreasing amounts of L-lysine-d-amphetamine or d-amphetamine in maleand female dogs.

FIG. 36. Left ventricular blood pressure following intravenous bolusinjection of increasing amounts of L-lysine-d-amphetamine ord-amphetamine in male and female dogs.

FIG. 37. Locomotor activity of rats following oral administration ofL-lysine-d-amphetamine or d-amphetamine (5 hour time-course).

FIG. 38. Locomotor activity of rats following oral administration ofL-lysine-d-amphetamine or d-amphetamine (12 hour time-course).

FIG. 39. Locomotor activity of rats following intranasal administrationof L-lysine-d-amphetamine or d-amphetamine (1 hour time-course).

FIG. 40. Locomotor activity of rats following intranasal administration(with carboxymethylcellulose) of L-lysine-d-amphetamine or d-amphetamine(2 hour time-course).

FIG. 41. Locomotor activity of rats following intravenous administrationof L-lysine-d-amphetamine or d-amphetamine (3 hour time-course).

FIG. 42. Intranasal bioavailability of abuse-resistant amphetamine aminoacid-, di-, and tri-peptide conjugates (ELISA analysis).

FIG. 43. Oral bioavailability of abuse-resistant amphetamine aminoacid-, di-, and tri-peptide conjugates (ELISA analysis).

FIG. 44. Intravenous bioavailability of an abuse-resistant amphetaminetri-peptide conjugate (ELISA analysis).

FIG. 45. Intranasal bioavailability of an abuse-resistant amphetamineamino acid conjugate (ELISA analysis).

FIG. 46. Oral bioavailability of an abuse-resistant amphetamine aminoacid conjugate (ELISA analysis).

FIG. 47. Intravenous bioavailability of abuse-resistant amphetamineamino acid-, di-, and tri-peptide conjugates (ELISA analysis).

FIG. 48. Intranasal bioavailability of an abuse-resistant amphetamineamino tri-peptide conjugate (ELISA analysis).

FIG. 49. Intranasal bioavailability of abuse-resistant amphetamine aminoacid-, and di-peptide conjugates (ELISA analysis).

FIG. 50. Intranasal bioavailability of an abuse-resistant amphetaminedi-peptide conjugate containing D- and L-amino acid isomers (ELISAanalysis).

FIGS. 51A-B. Plasma concentrations of d-amphetamine andL-lysine-d-amphetamine in ng/mL for the serum levels (FIG. 51A), and inng/g for brain tissue (FIG. 51B), following oral administration ofL-lysine-d-amphetamine or d-amphetamine sulfate (5 mg/kg d-amphetaminebase) to rats. Serum and brain tissue d-amphetamine andL-lysine-d-amphetamine concentrations were measured by LC/MS/MS(compound indicated in parenthesis).

FIGS. 52A-B. Plasma d-amphetamine and L-lysine-d-amphetamine levels(52A, ng/mL; 52B, μM) over a 72 hour period following oraladministration of L-lysine-d-amphetamine (25 mg L-lysine-d-amphetaminemesylate containing 7.37 mg d-amphetamine base) to humans (LC/MS/MSanalysis).

FIGS. 53A-B. Plasma d-amphetamine and L-lysine-d-amphetamine levels(53A, ng/mL; 53B, μM) over a 72 hour period following oraladministration of L-lysine-d-amphetamine (25 mg L-lysine-d-amphetaminemesylate containing 22.1 mg d-amphetamine base) to humans (LC/MS/MSanalysis).

FIGS. 54A-B. Plasma d-amphetamine levels (54A, 0-12 hours; 54B, 0-72hours) following oral administration of L-lysine-d-amphetamine (75 mgL-lysine-d-amphetamine mesylate containing 22.1 mg d-amphetamine base)or Adderall XR® (35 mg containing 21.9 mg amphetamine base to humans(LC/MS/MS analysis).

FIGS. 55A-B. Plasma d-amphetamine levels (55A, 0-12 hours; 55B, 0-72hours) following oral administration of L-lysine-d-amphetamine (75 mgL-lysine-d-amphetamine mesylate containing 22.1 mg d-amphetamine base)or Dexadrine Spansule® (30 mg containing 22.1 mg amphetamine base) tohumans (LC/MS/MS analysis).

DETAILED DESCRIPTION OF THE INVENTION

In accordance with the present invention and as used herein, thefollowing terms are defined with the following meanings, unlessexplicitly stated otherwise. For additional methods of attachingamphetamine to carriers, see application number U.S. Ser. No.10/156,527, and/or PCT/US03/05524 and/or PCT/US03/05525 each of which ishereby incorporated by reference in its entirety.

The invention utilizes covalent modification of amphetamine to decreaseits potential for causing overdose or abuse. The amphetamine iscovalently modified in a manner that decreases its pharmacologicalactivity, as compared to the unmodified amphetamine, at doses abovethose considered therapeutic. When given at lower doses, such as thoseintended for therapy, the covalently modified amphetamine retainspharmacological activity similar to that of the unmodified amphetamine.The covalent modification of amphetamine may comprise the attachment ofany chemical moiety through conventional chemistry.

Compounds, compositions and methods of the invention provide reducedpotential for overdose, reduced potential for abuse or addiction, and/orimprove amphetamine's characteristics with regard to high toxicities orsuboptimal release profiles. Without wishing to be limited to the belowtheory, we believe that overdose protection results from a naturalgating mechanism at the site of hydrolysis that limits the release ofthe active amphetamine from the prodrug at greater than therapeuticallyprescribed amounts. Therefore, abuse resistance is provided by limitingthe “rush” or “high” available from the active amphetamine released bythe prodrug and limiting the effectiveness of alternative routes ofadministration. Further, it is believed that the prodrug itself does notcross the blood brain barrier and is thus substantially absent from thecentral nervous system.

Throughout this application the use of “peptide” is meant to include asingle amino acid, a dipeptide, a tripeptide, an oligopeptide, apolypeptide, or the carrier peptide. Oligopeptide is meant to includefrom 2 amino acids to 70 amino acids. Further, at times the invention isdescribed as being an active agent attached to an amino acid, adipeptide, a tripeptide, an oligopeptide, polypeptide or carrier peptideto illustrate specific embodiments for the active agent conjugate.Preferred lengths of the conjugates and other preferred embodiments aredescribed herein.

Throughout this application the use of “chemical moiety” is meant toinclude at least amino acid(s), peptide(s), glycopeptide(s),carbohydrate(s), lipid(s), nucleoside(s), or vitamin(s).

Carbohydrates include sugars, starches, cellulose, and relatedcompounds. e.g., (CH₂O)_(n), wherein n is an integer larger than 2 orC_(n)(H₂O)_(n-1), with n larger than 5. More specific examples include,for instance, fructose, glucose, lactose, maltose, sucrose,glyceraldehyde, dihydroxyacetone, erythrose, ribose, ribulose, xylulose,galactose, mannose, sedoheptulose, neuraminic acid, dextrin, andglycogen.

A glycoprotein is a carbohydrate (or glycan) covalently linked toprotein. The carbohydrate may be in the form of a monosaccharide,disaccharide(s), oligosaccharide(s), polysaccharide(s), or theirderivatives (e.g. sulfo- or phospho-substituted).

A glycopeptide is a carbohydrate linked to an oligopeptide composed ofL- and/or D-amino acids. A glyco-amino-acid is a saccharide attached toa single amino acid by any kind of covalent bond. A glycosyl-amino-acidis a compound consisting of saccharide linked through a glycosyl linkage(O-, N- or S-) to an amino acid.

A “composition” as used herein refers broadly to any compositioncontaining a described molecule conjugate(s). The composition maycomprise a dry formulation, an aqueous solution, or a sterilecomposition. Compositions comprising the molecules described herein maybe stored in freeze-dried form and may be associated with a stabilizingagent such as a carbohydrate. In use, the composition may be deployed inan aqueous solution containing salts, e.g., NaCl, detergents, e.g.,sodium dodecyl sulfate (SDS), and other components.

“Amphetamine” shall mean any of the sympathomimetic phenethylaminederivatives which have central nervous system stimulant activity, suchas but not limited to, amphetamine, methamphetamine,p-methoxyamphetamine, methylenedioxyamphetamine,2,5-dimethoxy-4-methylamphetamine, 2,4,5-trimethoxyamphetamine and3,4-methylenedioxymethamphetamine.

Other embodiments are described according to the followingabbreviations.

-   -   Lys-Amp=L-lysine-d-amphetamine, Lys-Amph, Lysine-Amphetamine,        KAMP, K-amphetamine, or 2,6-diaminohexanoic        acid-(1-methyl-2-phenylethyl)-amide        Phe-Amp=Phenylalanine-Amphetamine, FAMP, or        2-amino-3-phenylpropanoic acid-(1-methyl-2-phenylethyl)-amide,        Ser-Amp=Serine-Amphetamine, SAMP, or 2-amino-3-hydroxylpropanoic        acid-(1-methyl-2-phenylethyl)-amide, Gly₃-Amp=GGG-Amphetamine,        GGGAMP, or        2-Amino-N-({[(1-methyl-2-phenyl-ethylcarbomyl)-methyl]-carbomyl}-methyl)-acetamide

This patent is meant to cover all compounds discussed regardless ofabsolute configurations. Thus, natural, L-amino acids are discussed butthe use of D-amino acids are also included. Similarly, references toamphetamine should be interpreted as inclusive of dextro- andlevo-isomers.

Furthermore, the following abbreviations may be used throughout thepatent.

-   -   BOC=t-butyloxycarbonyl    -   CMC=carboxymethylcellulose    -   DIPEA=di-isopropyl ethyl amine    -   mp=melting point    -   NMR=nuclear magnetic resonance    -   OSu=hydroxysuccinimido ester

“In a manner inconsistent with the manufacturer's instructions” is meantto include but is not limited to consuming amounts greater than amountsdescribed on the label or ordered by a licensed physician, and/oraltering by any means (e.g. crushing, breaking, melting, separatingetc.) the dosage formulation such that the composition maybe injected,inhaled or smoked.

Use of the phrases such as, “decreased”, “reduced”, “diminished” or“lowered” is meant to include at least a 10% change in pharmacologicalactivity with greater percentage changes being preferred for reductionin abuse potential and overdose potential. For instance, the change mayalso be greater than 25%, 35%, 45%, 55%, 65%, 75%, 85%, 95%, 96%, 97%,98%, 99%, or increments therein.

For each of the recited embodiments, the amphetamine may be any of theabove discussed stimulants. In one embodiment, the amphetamine isdextroamphetamine or methylphenidate.

The attached chemical moiety may be any chemical substance thatdecreases the pharmacological activity until amphetamine is released.Preferably the chemical moiety is a single amino acid, dipeptide ortripeptide. Amphetamine binds to specific sites to produce variouseffects (Hoebel, et al., 1989). The attachment of certain chemicalmoieties can therefore diminish or prevent binding to these biologicaltarget sites. Further, the covalent modification may prevent stimulantactivity by preventing the drug from crossing the blood-brain barrier.Preferably, absorption of the composition into the brain is prevented orsubstantially diminished and/or delayed when delivered by routes otherthan oral administration.

The attached chemical moiety may further comprise naturally occurring orsynthetic substances. This includes, but is not limited to, theattachment of amphetamine to amino acids, peptides, lipids,carbohydrates, glycopeptides, nucleic acids or vitamins. These chemicalmoieties could be expected to affect delayed release in thegastrointestinal tract and prevent rapid onset of the desired activity,particularly when delivered by parenteral routes. (Hoebel, B. G., L.Hernandez, et al., “Microdialysis studies of brain norepinephrine,serotonin, and dopamine release during ingestive behavior. Theoreticaland clinical implications.” Ann N Y Acad Sci 575: 171-91) (1989).

For each of the recited embodiments, the amino acid or peptide maycomprise of one or more of the naturally occurring (L-) amino acids:alanine, arginine, asparagine, aspartic acid, cysteine, glycine,glutamic acid, glutamine, histidine, isoleucine, leucine, lysine,methionine, proline, phenylalanine, serine, tryptophan, threonine,tyrosine, and valine. In another embodiment, the amino acid or peptideis comprised of one or more of the naturally occurring (D) amino acids:alanine, arginine, asparagine, aspartic acid, cysteine, glycine,glutamic acid, glutamine, histidine, isoleucine, leucine, lysine,methionine, proline, phenylalanine, serine, tryptophan, threonine,tyrosine, and valine. In another embodiment, the amino acid or peptideis comprised of one or more unnatural, non-standard or synthetic aminoacids such as, aminohexanoic acid, biphenylalanine, cyclohexylalanine,cyclohexylglycine, diethylglycine, dipropylglycine,2,3-diaminoproprionic acid, homophenylalanine, homoserine, homotyrosine,naphthylalanine, norleucine, ornithine, phenylalanine(4-fluoro),phenylalanine(2,3,4,5,6 pentafluoro), phenylalanine(4-nitro),phenylglycine, pipecolic acid, sarcosine,tetrahydroisoquinoline-3-carboxylic acid, and tert-leucine. In anotherembodiment, the amino acid or peptide comprises of one or more aminoacid alcohols, for example, serine and threonine. In another embodimentthe amino acid or peptide comprises of one or more N-methyl amino acids,for example, N-methyl aspartic acid.

In another embodiment, the specific carriers are utilized as a baseshort chain amino acid sequence and additional amino acids are added tothe terminus or side chain. In another embodiment, the above amino acidsequence may have one more of the amino acids substituted with one ofthe 20 naturally occurring amino acids. It is preferred that thesubstitution be with an amino acid which is similar in structure orcharge compared to the amino acid in the sequence. For instance,isoleucine (IIe)[I] is structurally very similar to leucine (Leu)[L],whereas, tyrosine (Tyr)[Y] is similar to phenylalanine (Phe)[F], whereasserine (Ser)[S] is similar to threonine (Thr)[T], whereas cysteine(Cys)[C] is similar to methionine (Met)[M], whereas alanine (Ala)[A] issimilar to valine (Val)[V], whereas lysine (Lys)[K] is similar toarginine (Arg)[R], whereas asparagine (Asn)[N] is similar to glutamine(Gln)[Q], whereas aspartic acid (Asp)[D] is similar to glutamic acid(Glu)[E], whereas histidine (His)[H] is similar to proline (Pro)[P], andglycine (Gly)[G] is similar to tryptophan (Trp)[W]. In the alternative,the preferred amino acid substitutions may be selected according tohydrophilic properties (i.e., polarity) or other common characteristicsassociated with the 20 essential amino acids. While preferredembodiments utilize the 20 natural amino acids for their GRAScharacteristics, it is recognized that minor substitutions along theamino acid chain which do not effect the essential characteristics ofthe amino acid chain are also contemplated.

In one embodiment, the carrier range is between one to 12 chemicalmoieties with one to 8 moieties being preferred. In another embodiment,the number of chemical moieties is selected from 1, 2, 3, 4, 5, 6, or 7.In another embodiment, the molecular weight of the carrier portion ofthe conjugate is below about 2,500, more preferably below about 1,000,and most preferably below about 500 kD. In one embodiment, the chemicalmoiety is a single lysine. In another embodiment, the chemical moiety isa lysine bound to an additional chemical moiety.

Another embodiment of the invention is a composition for preventingoverdose comprising amphetamine which has been covalently bound to achemical moiety.

Another embodiment of the invention is a composition for safelydelivering amphetamine comprising a therapeutically effective amount ofsaid amphetamine which has been covalently bound to a chemical moietywherein said chemical moiety reduces the rate of absorption of theamphetamine as compared to delivering the unbound amphetamine.

Another embodiment of the invention is a composition for reducingamphetamine toxicity comprising amphetamine which has been covalentlybound to a chemical moiety wherein said chemical moiety increases therate of clearance when given at doses exceeding those within thetherapeutic range of said amphetamine.

Another embodiment of the invention is a composition for reducingamphetamine toxicity comprising amphetamine which has been covalentlybound to a chemical moiety wherein said chemical moiety provides a serumrelease curve which does not increase above amphetamine's toxicity levelwhen given at doses exceeding those within the therapeutic range ofamphetamine.

Another embodiment of the invention is a composition for reducingbioavailability of amphetamine comprising amphetamine covalently boundto a chemical moiety wherein said bound amphetamine maintains asteady-state serum release curve which provides a therapeuticallyeffective bioavailability but prevents spiking or increased blood serumconcentrations compared to unbound amphetamine when given at dosesexceeding those within the therapeutic range of amphetamine.

Another embodiment of the invention is a composition for preventing aC_(max) spike for amphetamine when taken by mean other than orally whilestill providing a therapeutically effective bioavailability curve iftaken orally comprising an amphetamine which has been covalently boundto a chemical moiety.

Another embodiment of the invention is a composition for preventing atoxic release profile in a patient comprising amphetamine covalentlybound to a chemical moiety wherein said bound amphetamine maintains asteady-state serum release curve which provides a therapeuticallyeffective bioavailability but prevents spiking or increase blood serumconcentrations compared to unbound amphetamine.

Another embodiment of the invention is a compound of Formula I:

A-X_(n)—Z_(m)

wherein A is an amphetamine as defined herein; X is a chemical moiety asdefined herein and n is between 1 and 50 and increments thereof; and Zis a further chemical moiety different from X which acts as an adjuvantand m is between 1 and 50 and increments thereof. In another embodiment,n is between 1 and 50, more preferably between 1 and 10, and m is 0.

Embodiments of the invention provide amphetamine compositions whichallow the amphetamine to be therapeutically effective when delivered atthe proper dosage but reduces the rate of absorption or extent ofbioavailability of the amphetamine when given at doses exceeding thosewithin the therapeutic range of amphetamine. Embodiments of theinvention also provide amphetamine compositions wherein the covalentlybound chemical moiety increases the rate of clearance of amphetaminewhen given at doses exceeding those within the therapeutic range of theamphetamine.

In another embodiment, the amphetamine compositions have substantiallylower toxicity compared to unbound amphetamine. In another embodiment,the amphetamine compositions reduce or eliminate the possibility ofoverdose by oral administration. In another embodiment, the amphetaminecompositions reduce or eliminate the possibility of overdose byintranasal administration. In another embodiment, the amphetaminecompositions reduce or eliminate the possibility of overdose byinjection. In another embodiment, the amphetamine compositions reduce oreliminate the possibility of overdose by inhalation.

In another embodiment, the amphetamine conjugates of the invention mayfurther comprise a polymer blend which comprises a hydrophilic polymerand/or a water-insoluble polymer. The polymers may be used according toindustry standards to further enhance the sustained release/abuseresistant properties of the amphetamine conjugate without reducing theabuse resistance. For instance, a composition might include: about 70%to about 100% amphetamine conjugate by weight, from about 0.01% to about10% of a hydrophilic polymer (e.g. hydroxypropyl methylcellulose), fromabout 0.01% to about 2.5% of a water-insoluble polymer (e.g. acrylicresin), from about 0.01% to about 1.5% of additives (e.g. magnesiumstearate), and from about 0.01% to about 1% colorant by weight.

Hydrophilic polymers suitable for use in the sustained releaseformulations include one or more natural or partially or totallysynthetic hydrophilic gums such as acacia, gum tragacanth, locust beangum, guar gum, or karaya gum, modified cellulosic substances such asmethylcellulose, hydroxomethylcellulose, hydroxypropyl methylcellulose,hydroxypropyl cellulose, hydroxyethylcellulose, carboxymethylcellulose;proteinaceous substances such as agar, pectin, carrageen, and alginates;and other hydrophilic polymers such as carboxypolymethylene, gelatin,casein, zein, bentonite, magnesium aluminum silicate, polysaccharides,modified starch derivatives, and other hydrophilic polymers known tothose of skill in the art, or a combination of such polymers. Thesehydrophilic polymers gel and would dissolve slowly in aqueous acidicmedia thereby allowing the amphetamine conjugate to diffuse from the gelin the stomach. When the gel reaches the intestines it would dissolve incontrolled quantities in the higher pH medium to allow further sustainedrelease. Preferred hydrophilic polymers are the hydroxypropylmethylcelluloses such as those manufactured by The Dow Chemical Companyand known as Methocel ethers, such as Methocel E10M.

Other formulations may further comprise pharmaceutical additivesincluding, but not limited to: lubricants such as magnesium stearate,calcium stearate, zinc stearate, powdered stearic acid, hydrogenatedvegetable oils, talc, polyethylene glycol, and mineral oil; colorantssuch as Emerald Green Lake, FD&C Red No. 40, FD&C Yellow No. 6, D&CYellow No. 10, or FD&C Blue No. 1 and other various certified coloradditives (See 21 CFR, Part 74); binders such as sucrose, lactose,gelatin, starch paste, acacia, tragacanth, povidone polyethylene glycol,Pullulan and corn syrup; glidants such as colloidal silicon dioxide andtalc; surface active agents such as sodium lauryl sulfate, dioctylsodium sulfosuccinate, triethanolamine, polyoxyethylene sorbitan,poloxalkol, and quarternary ammonium salts; preservatives andstabilizers; excipients such as lactose, mannitol, glucose, fructose,xylose, galactose, sucrose, maltose, xylitol, sorbitol, chloride,sulfate and phosphate salts of potassium, sodium, and magnesium; and/orany other pharmaceutical additives known to those of skill in the art.In one preferred embodiment, a sustained release formulation furthercomprises magnesium stearate and Emerald Green Lake.

An amphetamine conjugate, which is further formulated with excipients,may be manufactured according to any appropriate method known to thoseof skill in the art of pharmaceutical manufacture. For instance, theamphetamine-conjugate and a hydrophilic polymer may be mixed in a mixerwith an aliquot of water to form a wet granulation. The granulation maybe dried to obtain hydrophilic polymer encapsulated granules ofamphetamine-conjugate. The resulting granulation may be milled,screened, then blended with various pharmaceutical additives such as,water insoluble polymers, and/or additional hydrophilic polymers. Theformulation may then tableted and may further be film coated with aprotective coating which rapidly dissolves or disperses in gastricjuices.

However, it should be noted that the amphetamine conjugate controls therelease of amphetamine into the digestive tract over an extended periodof time resulting in an improved profile when compared to immediaterelease combinations and prevention of abuse without the addition of theabove additives. In a preferred embodiment, no further sustained releaseadditives are required to achieve a blunted or reduced pharmacokineticcurve (e.g., reduced euphoric effect) while achieving therapeuticallyeffective amounts of amphetamine release when taken orally.

The compounds of the invention can be administered by a variety ofdosage forms. Any biologically-acceptable dosage form known to personsof ordinary skill in the art, and combinations thereof, arecontemplated. Examples of preferred dosage forms include, withoutlimitation, chewable tablets, quick dissolve tablets, effervescenttablets, reconstitutable powders, elixirs, liquids, solutions,suspensions, emulsions, tablets, multi-layer tablets, bi-layer tablets,capsules, soft gelatin capsules, hard gelatin capsules, caplets,lozenges, chewable lozenges, beads, powders, granules, particles,microparticles, dispersible granules, cachets and combinations thereof.

The most effective means for delivering the abuse-resistant compounds ofthe invention is orally, to permit maximum release of the amphetamine,and provide therapeutic effectiveness and/or sustained release whilemaintaining abuse resistance. When delivered by oral route theamphetamine is released into circulation, preferably over an extendedperiod of time as compared to amphetamine alone.

Formulations of the invention suitable for oral administration can bepresented as discrete units, such as capsules, caplets or tablets. Theseoral formulations also can comprise a solution or a suspension in anaqueous liquid or a non-aqueous liquid. The formulation can be anemulsion, such as an oil-in-water liquid emulsion or a water-in-oilliquid emulsion. The oils can be administered by adding the purified andsterilized liquids to a prepared enteral formula, which is then placedin the feeding tube of a patient who is unable to swallow.

Soft gel or soft gelatin capsules may be prepared, for example bydispersing the formulation in an appropriate vehicle (vegetable oils arecommonly used) to form a high viscosity mixture. This mixture is thenencapsulated with a gelatin based film using technology and machineryknown to those in the soft gel industry. The industrial units so formedare then dried to constant weight.

Chewable tablets, for example may be prepared by mixing the formulationswith excipients designed to form a relatively soft, flavored, tabletdosage form that is intended to be chewed rather than swallowed.Conventional tablet machinery and procedures, that is both directcompression and granulation, i.e., or slugging, before compression, canbe utilized. Those individuals involved in pharmaceutical solid dosageform production are versed in the processes and the machinery used asthe chewable dosage form is a very common dosage form in thepharmaceutical industry.

Film-coated tablets, for example may be prepared by coating tabletsusing techniques such as rotating pan coating methods or air suspensionmethods to deposit a contiguous film layer on a tablet.

Compressed tablets, for example may be prepared by mixing theformulation with excipients intended to add binding qualities todisintegration qualities. The mixture is either directly compressed orgranulated then compressed using methods and machinery known to those inthe industry. The resultant compressed tablet dosage units are thenpackaged according to market need, i.e., unit dose, rolls, bulk bottles,blister packs, etc.

The invention also contemplates the use of biologically-acceptablecarriers which may be prepared from a wide range of materials. Withoutbeing limited thereto, such materials include diluents, binders andadhesives, lubricants, plasticizers, disintegrants, colorants, bulkingsubstances, flavorings, sweeteners and miscellaneous materials such asbuffers and adsorbents in order to prepare a particular medicatedcomposition.

Binders may be selected from a wide range of materials such ashydroxypropylmethylcellulose, ethylcellulose, or other suitablecellulose derivatives, povidone, acrylic and methacrylic acidco-polymers, pharmaceutical glaze, gums, milk derivatives, such as whey,starches, and derivatives, as well as other conventional binders knownto persons skilled in the art. Exemplary non-limiting solvents arewater, ethanol, isopropyl alcohol, methylene chloride or mixtures andcombinations thereof. Exemplary non-limiting bulking substances includesugar, lactose, gelatin, starch, and silicon dioxide.

Preferred plasticizers may be selected from the group consisting ofdiethyl phthalate, diethyl sebacate, triethyl citrate, cronotic acid,propylene glycol, butyl phthalate, dibutyl sebacate, castor oil andmixtures thereof, without limitation. As is evident, the plasticizersmay be hydrophobic as well as hydrophilic in nature. Water-insolublehydrophobic substances, such as diethyl phthalate, diethyl sebacate andcastor oil are used to delay the release of water-soluble vitamins, suchas vitamin B6 and vitamin C. In contrast, hydrophilic plasticizers areused when water-insoluble vitamins are employed which aid in dissolvingthe encapsulated film, making channels in the surface, which aid innutritional composition release.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations of this invention can include othersuitable agents such as flavoring agents, preservatives andantioxidants. Such antioxidants would be food acceptable and couldinclude vitamin E, carotene, BHT or other antioxidants known to those ofskill in the art.

Other compounds which may be included by admixture are, for example,medically inert ingredients, e.g., solid and liquid diluent, such aslactose, dextrose, saccharose, cellulose, starch or calcium phosphatefor tablets or capsules, olive oil or ethyl oleate for soft capsules andwater or vegetable oil for suspensions or emulsions; lubricating agentssuch as silica, talc, stearic acid, magnesium or calcium stearate and/orpolyethylene glycols; gelling agents such as colloidal clays; thickeningagents such as gum tragacanth or sodium alginate, binding agents such asstarches, arabic gums, gelatin, methylcellulose, carboxymethylcelluloseor polyvinylpyrrolidone; disintegrating agents such as starch, alginicacid, alginates or sodium starch glycolate; effervescing mixtures;dyestuff; sweeteners; wetting agents such as lecithin, polysorbates orlaurylsulphates; and other therapeutically acceptable accessoryingredients, such as humectants, preservatives, buffers andantioxidants, which are known additives for such formulations.

For oral administration, fine powders or granules containing diluting,dispersing and/or surface-active agents may be presented in a draught,in water or a syrup, in capsules or sachets in the dry state, in anon-aqueous suspension wherein suspending agents may be included, or ina suspension in water or a syrup. Where desirable or necessary,flavoring, preserving, suspending, thickening or emulsifying agents canbe included.

Liquid dispersions for oral administration may be syrups, emulsions orsuspensions. The syrups may contain as carrier, for example, saccharoseor saccharose with glycerol and/or mannitol and/or sorbitol. Thesuspensions and the emulsions may contain a carrier, for example anatural gum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose or polyvinyl alcohol.

The dose range for adult human beings will depend on a number of factorsincluding the age, weight and condition of the patient. Tablets andother forms of presentation provided in discrete units convenientlycontain a daily dose, or an appropriate fraction thereof, of one or moreof the compounds of the invention. For example, units may contain from 5mg to 500 mg, but more usually from 10 mg to 250 mg, of one or more ofthe compounds of the invention.

It is also possible for the dosage form to combine any forms of releaseknown to persons of ordinary skill in the art. These include immediaterelease, extended release, pulse release, variable release, controlledrelease, timed release, sustained release, delayed release, long acting,and combinations thereof. The ability to obtain immediate release,extended release, pulse release, variable release, controlled release,timed release, sustained release, delayed release, long actingcharacteristics and combinations thereof is known in the art.

Compositions of the invention may be administered in a partial, i.e.,fractional dose, one or more times during a 24 hour period, a singledose during a 24 hour period of time, a double dose during a 24 hourperiod of time, or more than a double dose during a 24 hour period oftime. Fractional, double or other multiple doses may be takensimultaneously or at different times during the 24 hour period. Thedoses may be uneven doses with regard to one another or with regard tothe individual components at different administration times.

Likewise, the compositions of the invention may be provided in a blisterpack or other such pharmaceutical package. Further, the compositions ofthe present inventive subject matter may further include or beaccompanied by indicia allowing individuals to identify the compositionsas products for a prescribed treatment. The indicia may additionallyinclude an indication of the above specified time periods foradministering the compositions. For example, the indicia may be timeindicia indicating a specific or general time of day for administrationof the composition, or the indicia may be a day indicia indicating a dayof the week for administration of the composition. The blister pack orother combination package may also include a second pharmaceuticalproduct.

It will be appreciated that the pharmacological activity of thecompositions of the invention can be demonstrated using standardpharmacological models that are known in the art. Furthermore, it willbe appreciated that the inventive compositions can be incorporated orencapsulated in a suitable polymer matrix or membrane for site-specificdelivery, or can be functionalized with specific targeting agentscapable of effecting site specific delivery. These techniques, as wellas other drug delivery techniques, are well known in the art.

In another embodiment of the invention, the solubility and dissolutionrate of the composition is substantially changed under physiologicalconditions encountered in the intestine, at mucosal surfaces, or in thebloodstream. In another embodiment the solubility and dissolution ratesubstantially decrease the bioavailability of the amphetamine,particularly at doses above those intended for therapy. In anotherembodiment, the decrease in bioavailability occurs upon intranasaladministration. In another embodiment, the decrease in bioavailabilityoccurs upon intravenous administration.

For each of the described embodiments, one or more of the followingcharacteristics may be realized: The toxicity of the amphetamineconjugate is substantially lower than that of the unbound amphetamine.The covalently bound chemical moiety reduces or eliminates thepossibility of overdose by oral administration. The covalently boundchemical moiety reduces or eliminates the possibility of overdose orabuse by intranasal administration. The covalently bound chemical moietyreduces or eliminates the possibility of overdose or abuse by injection.

The invention further provides methods for altering amphetamines in amanner that decreases their potential for abuse. Methods of theinvention provide various ways to regulate pharmaceutical dosage throughcovalent attachment of amphetamine to different chemical moieties. Oneembodiment provides a method of preventing overdose comprisingadministering to an individual amphetamine which has been covalentlybound to a chemical moiety.

Another embodiment provides a method of safely delivering amphetaminecomprising providing a therapeutically effective amount of a amphetaminewhich has been covalently bound to a chemical moiety wherein thechemical moiety reduces the rate of absorption of amphetamine ascompared to delivering the unbound amphetamine.

Another embodiment provides a method of reducing amphetamine toxicitycomprising providing a patient with amphetamine which has beencovalently bound to a chemical moiety, wherein the chemical moietyincreases the rate of clearance of pharmacologically active amphetamine(i.e., released amphetamine) when given at doses exceeding those withinthe therapeutic range of amphetamine.

Another embodiment provides a method of reducing amphetamine toxicitycomprising providing a patient with amphetamine which has beencovalently bound to a chemical moiety, wherein the chemical moietyprovides a serum release curve which does not increase above theamphetamine's toxicity level when given at doses exceeding those withinthe therapeutic range for the unbound amphetamine.

Another embodiment provides a method of reducing bioavailability ofamphetamine comprising providing amphetamine covalently bound to achemical moiety, wherein the bound amphetamine maintains a steady-stateserum release curve which provides a therapeutically effectivebioavailability but prevents spiking or increase blood serumconcentrations compared to unbound amphetamine when given at dosesexceeding those within the therapeutic range for the unboundamphetamine.

Another embodiment provides a method of preventing a C_(max) spike foramphetamine while still providing a therapeutically effectivebioavailability curve comprising providing amphetamine which has beencovalently bound to a chemical moiety.

In another embodiment, methods of the invention provide bioavailabilitycurves similar to those of FIGS. 6-55.

Another embodiment provides a method for preventing a toxic releaseprofile in a patient comprising administering to a patient amphetaminecovalently bound to a chemical moiety, wherein said bound amphetaminemaintains a steady-state serum release curve which provides atherapeutically effective bioavailability but prevents spiking orincrease blood serum concentrations compared to unbound amphetamine,particularly when taken at doses above prescribed amounts.

Another embodiment of the invention is a method for reducing orpreventing abuse of amphetamine comprising providing, administering, orprescribing said composition to a human in need thereof, wherein saidcomposition comprises a chemical moiety covalently attached toamphetamine such that the pharmacological activity of amphetamine isdecreased when the composition is used in a manner inconsistent with themanufacturer's instructions.

Another embodiment of the invention is a method for reducing orpreventing abuse of amphetamine comprising consuming an amphetamineconjugate of the invention, wherein said conjugate comprises a chemicalmoiety covalently attached to amphetamine such that the pharmacologicalactivity of amphetamine is substantially decreased when the compositionis used in a manner inconsistent with the manufacturer's instructions.

Another embodiment of the invention is a method of preventing overdoseof amphetamine comprising providing, administering, or prescribing anamphetamine composition of the invention to a human in need thereof,wherein said composition comprises a chemical moiety covalently attachedto amphetamine in a manner that decreases the potential of overdose fromamphetamine.

Another embodiment of the invention is a method of preventing overdoseof amphetamine, comprising consuming an amphetamine composition of theinvention, wherein said composition comprises a chemical moietycovalently attached to amphetamine in a manner that decreases thepotential of overdose from amphetamine.

Another embodiment of the invention is a method for reducing orpreventing the euphoric effect of amphetamine comprising providing,administering, or prescribing said to a human in need thereof, acomposition comprising a chemical moiety covalently attached toamphetamine such that the pharmacological activity of amphetamine isdecreased when the composition is used in a manner inconsistent with themanufacturer's instructions.

Another embodiment of the invention is a method for reducing orpreventing the euphoric effect of amphetamine, comprising consuming asaid composition comprising a chemical moiety covalently attached toamphetamine such that the pharmacological activity of amphetamine isdecreased when the composition is used in a manner inconsistent with themanufacturer's instructions.

Another embodiment of the invention is any of the preceding methodswherein said amphetamine composition is adapted for oral administration,and wherein said amphetamine is resistant to release from said chemicalmoiety when the composition is administered parenterally, such asintranasally or intravenously. Preferably, said amphetamine may bereleased from said chemical moiety in the presence of acid and/orenzymes present in the stomach, intestinal tract, or blood serum.Optionally, said composition may be in the form of a tablet, capsule,oral solution, oral suspension, or other oral dosage form discussedherein.

For each of the recited methods, the chemical moiety may be one or moreamino acid(s), oligopeptide(s), polypeptide(s), carbohydrate(s),glycopeptide(s), nucleic acid(s), or vitamin(s). Preferably, saidchemical moiety is an amino acid, oligopeptide, or polypeptide orcarbohydrate. Where the chemical moiety is a polypeptide, preferablysaid polypeptide comprises fewer than 70 amino acids, fewer than 50amino acids, fewer than 10 amino acids, or fewer than 4 amino acids.Where the chemical moiety is an amino acid, preferably said amino acidis lysine, serine, phenylalanine or glycine. Most preferably, said aminoacid is lysine.

For each of the recited embodiments, covalent attachment may comprise anester or carbonate bond.

For each of the recited methods, the composition may yield a therapeuticeffect without substantial euphoria. Preferably, said amphetaminecomposition provides a therapeutically bioequivalent AUC when comparedto amphetamine alone but does provide a C_(max) which results ineuphoria.

Another embodiment of the invention is a method for reducing orpreventing abuse of amphetamine comprising orally administering anamphetamine composition of the invention to a human in need thereof,wherein said composition comprises an amino acid or peptide (e.g.,lysine) covalently attached to amphetamine such that the pharmacologicalactivity of amphetamine is decreased when the composition is used in amanner inconsistent with the manufacturer's instructions.

Another embodiment is a method of preventing overdose of a amphetaminecomprising orally administering an amphetamine composition to a human inneed thereof, wherein said composition comprises an amino acid orpeptide (e.g., lysine) covalently attached to amphetamine in a mannerthat decreases the potential of amphetamine to result in overdose.

Another embodiment is a method for reducing or preventing the euphoriceffect of amphetamine comprising orally administering an amphetaminecomposition to a human in need thereof, wherein said compositioncomprises an amino acid or peptide (e.g., lysine) covalently attached toamphetamine such that the pharmacological activity of amphetamine isdecreased when the composition is used in a manner inconsistent with themanufacturer's instructions.

For each of the recited methods of the invention the followingproperties may be achieved through bonding amphetamine to the chemicalmoiety. In one embodiment, the toxicity of the compound may be lowerthan that of the amphetamine when delivered in its unbound state or as asalt thereof. In another embodiment, the possibility of overdose by oraladministration is reduced or eliminated. In another embodiment, thepossibility of overdose by intranasal administration is reduced oreliminated. In another embodiment, the possibility of overdose byinjection administration is reduced or eliminated.

Another embodiment of the invention provides methods of treating variousdiseases or conditions comprising administering compounds orcompositions of the invention which further comprise commonly prescribedactive agents for the respective illness or diseases wherein theamphetamine is covalently attached to a chemical moiety. For instance,one embodiment of the invention comprises a method of treating attentiondeficit hyperactivity disorder (ADHD) comprising administering to apatient amphetamine covalently bound to a chemical moiety. Anotherembodiment provides a method of treating attention deficit disorder(ADD) comprising administering to a patient compounds or compositions ofthe invention, amphetamine covalently bound to a chemical moiety.

Another embodiment of the invention provides a method of treatingnarcolepsy comprising administering to a patient compounds orcompositions of the invention.

In order to facilitate a more complete understanding of the invention.Examples are provided below. However, the scope of the invention is notlimited to specific embodiments disclosed in these Examples, which arefor purposes of illustration only.

Examples Example 1 General Synthesis of Amino Acid-AmphetamineConjugates

Amino acid conjugates were synthesized by the general method describedin FIGS. 1-5.

Example 2 Synthesis of L-lysine-d-amphetamine

L-lysine-d-amphetamine was synthesized (see FIG. 2) by the followingmethod:

a. Coupling Reagents MW Weight mmoles Molar Equivalents d-amphetamine135.2 4.75 g 35.13 1 freebase Boc-Lys(Boc)-OSu 443.5 15.58 g 35.13 1Di-iPr-Et-Amine 129 906 mg 7.03 0.2, d = 0.74, 1.22 mL 1,4-Dioxane — 100mL — —

To a solution of Boc-Lys(Boc)-OSu (15.58 g, 35.13 mmol) in dioxane (100mL) under an inert atmosphere was added d-amphetamine freebase (4.75 g,35.13 mmol) and DiPEA (0.9 g, 1.22 mL, 7.03 mmol). The resulting mixturewas allowed to stir at room temperature overnight. Solvent and excessbase were then removed using reduced pressure evaporation. The crudeproduct was dissolved in ethyl acetate and loaded on to a flash column(7 cm wide, filled to 24 cm with silica) and eluted with ethyl acetate.The product was isolated; the solvent reduced by rotary evaporation andthe purified protected amide was dried by high-vac to obtain a whitesolid. ¹H NMR (DMSO-d₆) δ 1.02-1.11 (m, 2H, Lys γ-CH₂), δ 1.04 (d, 3H,Amp α-CH₃), δ 1.22-1.43 (m, 4H, Lys-β and δ-CH₂), δ 1.37 (18H, Boc, 6×CH₃), δ 2.60-2.72 (2H, Amp CH₂), δ 3.75-3.83, (m, 1H, Lys α-H) δ 3.9-4.1(m, 1H, Amp α-H), δ 6.54-6.61 (d, 1H, amide NH), δ 6.7-6.77 (m, 1H,amide NH), δ 7.12-7.29 (m, 5H, ArH), δ 7.65-7.71 (m, 1, amide NH);mp=86-88° C.

b. Deprotection Molar Reagents MW Weight mmoles Equivalents 4M HCl indioxane 4 mmol/mL 50 mL 200 6.25 Boc-Lys(Boc)-Amp 463.6 14.84 g 32 11,4-Dioxane — 50 mL — —

The protected amide was dissolved in 50 mL of anhydrous dioxane andstirred while 50 mL (200 mmol) of 4M HCl/dioxane was added and stirredat room temperature overnight. The solvents were then reduced by rotaryevaporation to afford a viscous oil. Addition of 100 mL MeOH followed byrotary evaporation resulted in a golden colored solid material that wasfurther dried by storage at room temperature under high vacuum. ¹H NMR(DMSO-d₆) δ 0.86-1.16 (m, 2H, Lys γ-CH₂), δ 1.1 (d, 3H, Amp α-CH₃), δ1.40-1.56 (m, 4H, Lys-β and δ-CH₂), δ 2.54-2.78 (m, 2H, Amp CH₂, 2H, Lysε-CH₂), 3.63-3.74 (m, 1H, Lys α-H), δ 4.00-4.08 (m, 1H, Amp α-H), δ7.12-7.31 (m, 5H, Amp ArH), δ 8.13-8.33 (d, 3H, Lys amine) δ 8.70-8.78(d, 1H, amide NH); mp=120-122° C.

Example 3 Synthesis of Ser-Amp

Ser-Amp was synthesized by a similar method (see FIG. 3) except theamino acid starting material was Boc-Ser(O-tBu)-OSu and the deprotectionwas done using a solution of trifluoroacetic acid instead of HCl.

Example 4 Synthesis of Phe-Amp

Phe-Amp was synthesized by a similar method (see FIG. 4) except theamino acid starting material was Boc-Phe-OSu.

Example 5 Synthesis of Gly₃-Amp

Gly₃-Amp was synthesized by a similar method (see FIG. 5) except theamino acid starting material was Boc-GGG-OSu.

Example 6 Pharmacokinetics of L-lysine-d-amphetamine Compared tod-amphetamine Sulfate (ELISA Analysis)

Male Sprague-Dawley rats were provided water ad libitum, fastedovernight and dosed by oral gavage L-lysine-d-amphetamine ord-amphetamine sulfate. In all studies doses contained equivalent amountsof d-amphetamine base. Plasma d-amphetamine concentrations were measuredby ELISA (Amphetamine Ultra, 109319, Neogen, Corporation, Lexington,Ky.). The assay is specific for d-amphetamine with only minimalreactivity (0.6%) of the major d-amphetamine metabolite(para-hydroxy-d-amphetamine) occurring. L-lysine-d-amphetamine was alsodetermined to be essentially unreactive in the ELISA (<1%).

Mean (n=4) plasma concentration curves of d-amphetamine orL-lysine-d-amphetamine are shown in FIG. 6. Extended release wasobserved in all four L-lysine-d-amphetamine dosed animals and C_(max)was substantially decreased as compared to animals dosed withd-amphetamine sulfate. Plasma d-amphetamine concentrations of individualanimals for d-amphetamine or L-lysine-d-amphetamine are shown inTable 1. The mean plasma d-amphetamine concentrations are shown in Table2. The time to peak concentration for L-lysine-d-amphetamine was similarto that of d-amphetamine. Pharmacokinetic parameters for oraladministration of d-amphetamine or L-lysine-d-amphetamine are summarizedin Table 3.

TABLE 1 Plasma Concentrations of d-amphetamine from Individual AnimalsOrally Administered d-amphetamine or L-lysine-d-amphetamine (3 mg/kgd-amphetamine base). d-amphetamine (ng/ml) L-lysine-d-amphetamine(ng/ml) Time Rat Rat Rat Rat Rat (hours) #1 #2 #3 #4 #1 Rat #2 Rat #3Rat #4 0.5 144 157 101 115 52 62 74 44 1 152 78 115 78 48 72 79 57 1.585 97 117 95 42 62 76 53 3 34 45 72 38 61 60 71 43 5 20 14 12 15 49 3344 22 8 3 3 2 2 15 14 12 8

TABLE 2 Mean Plasma Concentrations of d-amphetamine Following OralAdministration of d-amphetamine or L-lysine-d-amphetamine. Plasmad-amphetamine Concentrations (ng/ml) L-lysine-d- d-amphetamineamphetamine Hours Mean +/−SD CV Mean +/−SD CV 0.5 129 25 20 58 13 22 1106 35 33 64 14 22 1.5 99 13 14 58 14 25 3 47 17 36 59 11 19 5 15 4 2437 12 32 8 2 1 35 12 3 24

TABLE 3 Pharmacokinetic Parameters of d-amphetamine Following OralAdministration of d-amphetamine or L-lysine-d-amphetamine. AUC (0-8 h)Percent Cmax Percent Mean Peak Percent Drug ng/ml h Amphetamine (ng/ml)Amphetamine (ng/ml) Amphetamine Amphetamine 341 +/− 35 100 111 +/− 27100 129 100 Lys-Amp 333 +/− 66 98  61 +/− 13 55 64 50

Example 6 illustrates that when lysine is conjugated to the active agentamphetamine the peak levels of amphetamine are decreased whilebioavailability is maintained approximately equal to amphetamine. Thebioavailability of amphetamine released from L-lysine-d-amphetamine issimilar to that of amphetamine sulfate at the equivalent dose, thusL-lysine-d-amphetamine maintains its therapeutic value. The gradualrelease of amphetamine from L-lysine-d-amphetamine and decrease in peaklevels reduce the possibility of overdose.

Example 7 Oral Bioavailability of L-lysine-d-amphetamine at VariousDoses Approximating a Range of Therapeutic Human Doses

Mean (n=4) plasma concentration curves of d-amphetamine vs.L-lysine-d-amphetamine are shown for rats orally administered 1.5, 3,and 6 mg/kg in FIGS. 7, 8 and 9, respectively. Extended release wasobserved at all three doses for L-lysine-d-amphetamine dosed animals.The mean plasma concentrations for 1.5, 3, and 6 mg/kg are shown inTables 4, 5 and 6, respectively. Pharmacokinetic parameters for oraladministration of d-amphetamine vs. L-lysine-d-amphetamine at thevarious doses are summarized in Table 7.

TABLE 4 Mean Plasma Concentrations of d-amphetamine vs. L-lysine-d-amphetamine Following Oral Admistration (1.5 mg/kg) Plasma AmphetamineConcentrations (ng/ml) L-lysine-d- d-amphetamine amphetamine Hours Mean+/−SD CV Mean +/−SD CV 0 0 0 0 0 0 0 0.25 103 22 21 31 11 37 0.5 126 2016 51 23 45 1 101 27 27 68 23 34 1.5 116 28 24 72 10 14 3 66 13 20 91 55 5 40 7 18 75 16 22 8 17 2 15 39 13 34

TABLE 5 Mean Plasma Concentrations of d-amphetamine vs. L-lysine-d-amphetamine Following Oral Admistration (3 mg/kg) Plasma AmphetamineConcentrations (ng/ml) L-lysine-d- d-amphetamine amphetamine Hours Mean+/−SD CV Mean +/−SD CV 0 0 0 0.25 96 41 43 51 49 97 0.5 107 49 46 36 3596 1 121 17 14 81 44 54 1.5 120 33 27 97 32 33 3 91 30 33 88 13 15 5 6222 36 91 21 23 8 19 6 33 46 16 34

TABLE 6 Mean Plasma Concentrations of d-amphetamine vs. L-lysine-d-amphetamine Following Oral Admistration (6 mg/kg). Plasma AmphetamineConcentrations (ng/ml) L-lysine-d- d-amphetamine amphetamine Hours Mean+/−SD CV Mean +/−SD CV 0 0 0 0.25 204 14 7 74 38 51 0.5 186 9 5 106 3937 1 167 12 7 133 33 24 1.5 161 24 15 152 22 15 3 111 29 26 157 15 10 578 9 11 134 18 13 8 35 5 15 79 12 15

TABLE 7 Pharmacokinetic Parameters of d-amphetamine Following OralAdministration of d-amphetamine or L-lysine-d-amphetamine. 1.5 mg/kg 3mg/kg 6 mg/kg L-lysine-d- L-lysine-d- L-lysine-d- Parameterd-amphetamine amphetamine d-amphetamine amphetamine d-amphetamineamphetamine AUC (ng/ml 481 538 587 614 807 1005 h) Percent 100 112 100105 100 125 Cmax 133 93 587 614 807 1005 (ng/ml) Percent 100 70 100 105100 125 Tmax 0.938 3.5 1 1.56 0.563 2.625 (hours) Percent 100 373 100156 100 466

Example 8 Oral Bioavailability of L-lysine-d-amphetamine at VariousDoses Approximating a Range of Therapeutic Human Doses Compared to aSuprapharmacological Dose

Male Sprague-Dawley rats were provided water ad libitum, fastedovernight and dosed by oral gavage with 1.5, 3, 6, 12, and 60 mg/kg ofamphetamine sulfate or L-lysine-d-amphetamine containing the equivalentamounts of d-amphetamine. Concentrations of d-amphetamine were measuredby ELISA.

It has been demonstrated that when lysine is conjugated to the activeagent d-amphetamine the levels of d-amphetamine at 30 minutespost-administration are decreased by approximately 50% over a dose rangeof 1.5 to 12 mg/kg. However, when a suprapharmcological dose (60 mg/kg)is given the levels of d-amphetamine from L-lysine-d-amphetamine onlyreached 8% of those seen for d-amphetamine sulfate (Tables 8 and 9, FIG.10). The substantial decrease in oral bioavailability at a high dosegreatly reduces the abuse potential of L-lysine-d-amphetamine.

TABLE 8 Levels of d-amphetamine vs. Dosage at 0.5 h Post Dosing withd-amphetamine Sulfate. Dose mg/kg 1.5 3 6 12 60 ng/ml 0.5 h 109 +/− 59196 +/− 72 294 +/− 202 344 +/− 126 3239 +/− 73 Percent 100 100 100 100100

TABLE 9 Levels of d-amphetamine vs. Dosage at 0.5 h Post Dosing withL-lysine-d- amphetamine. Dose mg/kg 1.5 3 6 12 60 ng/ml 0.5 h 45 +/− 1086 +/− 26 129 +/− 46 172 +/− 113 266 +/− 18 Percent 41 44 44 50 8

Example 9 Decreased Oral Bioavailability of L-lysine-d-amphetamine at aHigh Dose

An additional oral PK study illustrated in FIG. 11 shows thed-amphetamine blood levels of a 60 mg/kg dose over an 8 h time course.In the case of d-amphetamine blood levels quickly reached a very highlevel and 8 of 12 animals either died or were sacrificed due to acutesymptoms of toxicity. Blood levels (Tables 10-11) of animalsadministered L-lysine-d-amphetamine, on the other hand, did not peakuntil 5 hours and reached only a fraction of the levels of the animalsreceiving amphetamine (note: valid data past 3 h for d-amphetamine couldnot be determined due to death and sacrifice of animals).

TABLE 10 Mean Plasma Concentrations of d-amphetamine vs. L-lysine-d-amphetamine Following Oral Administration of a High Dose (60 mg/kg).Plasma Amphetamine Concentrations (ng/ml) L-lysine-d- d-amphetamineamphetamine Hours Mean +/− SD CV Mean +/− SD CV 0 NA NA NA NA NA NA 0.252174 907 42 35 17 48 0.5 2643 578 22 81 33 41 1 2828 1319 47 212 30 141.5 2973 863 29 200 79 40 3 2944 95 3 440 133 30 5 NA NA NA 565 100 18 8NA NA NA 410 206 50

TABLE 11 Pharmacokinetic Parameters of d-amphetamine vs.L-lysine-d-amphetamine AUC Percent Cmax Percent Mean Peak Percent Drugng/ml h d-amphetamine (ng/ml) d-amphetamine (ng/ml) d-amphetamined-mphetamine 8,130 100 3623 100 2973 100 L-lysine-d- 3,143 39 582 16 56519 amphetamine

Example 10 Oral Bioavailability of d-amphetamine FollowingAdministration of an Extended Release Formulation (Intact or Crushed) orL-lysine-d-amphetamine

Doses of an extended release formulation of d-amphetamine sulfate(Dexadrine Spansule capsules) were orally administered to rats as intactcapsules or as crushed capsules and compared to a dose ofL-lysine-d-amphetamine containing an equivalent amount of d-amphetaminebase (FIG. 14). The crushed capsules showed an increase in C_(max) andAUC_(inf) of 84 and 13 percent, respectively, as compared to intactcapsules (Tables 12-13). In contrast, C_(max) and AUC_(inf) ofd-amphetamine following administration of L-lysine-d-amphetamine weresimilar to that of the intact capsule illustrating that extended releaseis inherent to the compound itself and can not be circumvented by simplemanipulation.

TABLE 12 Time-course Concentrations of d-amphetamine Following OralAdministration of Extended Release Dexadrine Spansule Capsules orCrushed Extended Release Dexadrine Spansule Capsules orL-lysine-d-amphetamine at Doses Containing 3 mg/kg d-Amphetamine Base.Plasma Concentration (ng/ml) Intact Spansule Crushed Spansule HoursCapsule Capsule L-lysine-d-amphetamine 0 0 0 0 0.25 32 46 3 0.5 33 85 51 80 147 34 1.5 61 101 60 3 64 66 76 5 46 39 66 8 34 12 38

TABLE 13 Time-course Concentrations of d-amphetamine Following OralAdministration of Extended Release Dexadrine Spansule Capsules orCrushed Extended Release Dexadrine Spansule Capsules orL-lysine-d-amphetamine at Doses Containing 3 mg/kg d-Amphetamine Base.Intact Spansule Crushed Spansule L-lysine- Parameter Capsule Capsuled-amphetamine AUC_(0-8 h) (ng · h/ml) 399 449 434 Percent 100 113 109C_(max) (ng/ml) 80 147 76 Percent 100 184 95 T_(max) (hours) 1 1 3Percent 100 100 300

Example 10 illustrates the advantage of the invention over conventionalcontrolled release formulations of d-amphetamine.

Example 11 Decreased Intranasal Bioavailability ofL-lysine-d-amphetamine vs. Amphetamine

Male Sprague-Dawley rats were dosed by intranasal administration with 3mg/kg of amphetamine sulfate or L-lysine-d-amphetamine hydrochloridecontaining the equivalent amounts of d-amphetamine.L-lysine-d-amphetamine did not release any significant amount ofd-amphetamine into circulation by IN administration. Mean (n=4) plasmaamphetamine concentration curves of amphetamine vs.L-lysine-d-amphetamine are shown in FIG. 12. Pharmacokinetic parametersfor IN administration of L-lysine-d-amphetamine are summarized in Table14.

TABLE 14 Pharmacokinetic Parameters of Amphetamine vs.L-lysine-d-amphetamine by IN Administration. AUC (0-1.5 h) Percent CmaxPercent Drug ng/ml h d-amphetamine (ng/ml) d-amphetamine Amphetamine 727100 1,377 100 L-lysine-d- 4 0.5 7 0.5 amphetamine

Example 11 illustrates that when lysine is conjugated to the activeagent d-amphetamine the bioavailability by the intranasal route issubstantially decreased thereby diminishing the ability to abuse thedrug by this route.

Example 12 Intravenous Bioavailability of Amphetamine vs.L-lysine-d-amphetamine

Male Sprague-Dawley rats were dosed by intravenous tail vein injectionwith 1.5 mg/kg of d-amphetamine or L-lysine-d-amphetamine containing theequivalent amount of amphetamine. As observed with IN dosing, theconjugate did not release a significant amount of d-amphetamine. Mean(n=4) plasma concentration curves of amphetamine vs.L-lysine-d-amphetamine are shown in FIG. 13. Pharmacokinetic parametersfor IV administration of L-lysine-d-amphetamine are summarized in Table15.

TABLE 15 Pharmacokinetic Parameters of d-amphetamine vs.L-lysine-d-amphetamine by IV Administration. AUC (0-1.5 h) % Cmax % Drugng/ml h Amphetamine (ng/ml) Amphetamine Amphetamine 190 100 169 100K-amphetamine 6 3 5 3

Example 12 illustrates that when lysine is conjugated to the activeagent amphetamine the bioavailability of amphetamine by the intravenousroute is substantially decreased, thereby diminishing the ability toabuse the drug by this route.

Example 13 Oral Bioavailability of L-lysine-d-amphetamine Compared tod-amphetamine at Escalating Doses

As shown in FIGS. 15-19, the fraction of intact L-lysine-d-amphetamineabsorbed following oral administration in rats increased non-linearly inproportion to escalating doses from 1.5 to 12 mg/kg (d-amphetaminebase). The fraction absorbed at 1.5 mg/kg was only 2.6 percent whereasit increased to 24.6 percent by 12 mg/kg. The fraction absorbed fell to9.3 percent at the high dose of 60 mg/kg. T_(max) ranged from 0.25 to 3hours and peak concentrations occurred earlier than for d-amphetamine inL-lysine-d-amphetamine dosed rats. L-lysine-d-amphetamine was clearedmore rapidly than d-amphetamine with nearly undetectable concentrationsby 8 hours at the lowest dose.

T_(max) for d-amphetamine from L-lysine-d-amphetamine ranged from 1.5 to5 hours as compared to 0.5 to 1.5 following administration ofd-amphetamine sulfate. The difference in time to reach maximumconcentration was greater at higher doses. C_(max) of d-amphetaminefollowing oral delivery of L-lysine-d-amphetamine was reduced byapproximately half as compared to C_(max) following d-amphetaminesulfate administration at doses of 1.5 to 6 mg/kg, approximating humanequivalent doses (HEDs) in the therapeutic range (HED d-amphetaminesulfate; 19.9 to 39.9 mg). HEDs are defined as the equivalent dose for a60 kg person in accordance to the body surface area of the animal model.The adjustment factor for rats is 6.2. The HED for a rat dose of 1.5mg/kg of d-amphetamine, for example, is equivalent to 1.5/6.2×60=14.52d-amphetamine base; which is equivalent to 14.52/0.7284=19.9 mgd-amphetamine sulfate, when adjusted for the salt content.

At doses above HEDs in the targeted therapeutic range (12 and 60 mg/kg;HED d-amphetamine sulfate 79.8 and 399 mg), C_(max) was reduced by 73and 84 percent, respectively, as compared to d-amphetamine sulfate. AUCsof d-amphetamine following oral administration of L-lysine-d-amphetaminewere similar to those of d-amphetamine sulfate at lower doses. Asobserved with C_(max), however, the AUCs for d-amphetamine fromL-lysine-d-amphetamine were substantially decreased compared to those ofd-amphetamine sulfate at higher doses with the AUC_(inf) reduced by 76%at the highest dose (60 mg/kg; HED 399 mg d-amphetamine sulfate.

In summary, oral bioavailability of d-amphetamine fromL-lysine-d-amphetamine decreased to some degree at higher doses in rats.However, pharmacokinetics with respect to dose were nearly linear forL-lysine-d-amphetamine at doses from 1.5 to 60 mg/kg (HED d-amphetaminesulfate; 19.9 to 797.2 mg) with the fraction absorbed ranging from 52 to81 percent (extrapolated form 1.5 mg/kg dose). Pharmacokinetics ofd-amphetamine sulfate was also nearly linear at lower doses of 1.5 to 6mg/kg (HED; 19.9 to 79.7) with the fraction absorbed ranging form 62 to84. In contrast to L-lysine-d-amphetamine, however, parameters weredisproportionately increased at higher doses for d-amphetamine sulfatewith the fraction absorbed calculated as 101 and 223 percent(extrapolated form 1.5 mg/kg dose), respectively, for thesuprapharmacological doses of 12 and 60 mg/kg (HED d-amphetaminesulfate; 159.4 and 797.2 mg).

The results suggest that the capacity for clearance of d-amphetaminewhen delivered as the sulfate salt becomes saturated at the higher doseswhereas the gradual hydrolysis of L-lysine-d-amphetamine precludessaturation of d-amphetamine elimination at higher doses. The differencein proportionality of dose to bioavailability (Cmax and AUC) ford-amphetamine and L-lysine-d-amphetamine is illustrated in FIGS. 20-22.The pharmacokinetic properties of L-lysine-d-amphetamine as compared tod-amphetamine at the higher doses decrease the ability to escalatedoses. This improves the safety and reduces the abuse liability ofL-lysine-d-amphetamine as a method of delivering d-amphetamine for thetreatment of ADHD or other indicated conditions.

Example 14 Intranasal Bioavailability of L-lysine-d-amphetamine Comparedto d-amphetamine

As shown in FIGS. 23-24, bioavailability of d-amphetamine followingbolus intranasal administration of L-lysine-d-amphetamine wasapproximately 5 percent of that of the equivalent d-amphetamine sulfatedose with AUC_(inf) values of 56 and 1032, respectively. C_(max) ofd-amphetamine following L-lysine-d-amphetamine administration by theintranasal route was also about 5 percent of that of the equivalentamount of d-amphetamine sulfate with values of 78.6 ng/mL and 1962.9ng/mL, respectively. As with intravenous administration, T_(max) ofd-amphetamine concentration was delayed substantially forL-lysine-d-amphetamine (60 minutes) as compared to T_(max) ofd-amphetamine sulfate (5 minutes), again reflecting the gradualhydrolysis of L-lysine-d-amphetamine. A high concentration of intactL-lysine-d-amphetamine was detected following intranasal dosingsuggesting that the large decrease in bioavailability of d-amphetaminewas due to minimal hydrolysis of L-lysine-d-amphetamine when deliveredby this route. It appears that only minimal amounts of d-amphetamine canbe delivered by intranasal administration of L-lysine-d-amphetamine.

Example 15 Intravenous Bioavailability of L-lysine-d-amphetamineCompared to d-amphetamine

As shown in FIGS. 25-26, bioavailability of d-amphetamine followingbolus intravenous administration of L-lysine-d-amphetamine wasapproximately one-half that of the equivalent d-amphetamine sulfate dosewith AUC_(inf) values of 237.8 and 420.2, respectively. C_(max) ofd-amphetamine following L-lysine-d-amphetamine administration was onlyabout one-fourth that of the equivalent amount of d-amphetamine withvalues of 99.5 and 420.2, respectively. T_(max) of d-amphetamineconcentration was delayed substantially for L-lysine-d-amphetamine (30minutes) as compared to T_(max) of d-amphetamine sulfate (5 minutes),reflecting the gradual hydrolysis of L-lysine-d-amphetamine. Inconclusion, the bioavailability of d-amphetamine by the intravenousroute is substantially decreased and delayed when given asL-lysine-d-amphetamine. Moreover, bioavailability is less than thatobtained by oral administration of the equivalent dose ofL-lysine-d-amphetamine.

Summary of LC/MS/MS Bioavailability Data in Rats

The following tables summarize the bioavailability data collected in theexperiments discussed in examples 13-15. Tables 15-17 summarize thepharmacokinetic parameters of d-amphetamine following oral, intranasal,or bolus intravenous administration of d-amphetamine orL-lysine-d-amphetamine.

TABLE 15 Pharmacokinetic Parameters of d-amphetamine Following OralAdministration of L-lysine-d-amphetamine or d-amphetamine at EscalatingDoses. Dose Cmax Tmax AUC(0-8) AUC(inf) F AUC/Dose Cmax/Dose Route Drug(mg/kg) (ng/mL) (h) (ng · mL/h) (ng · mL/h) (%) (ng · h · kg/mL/mg) ng ·kg/mL/mg Oral L-lysine- 1.5 59.6 3 308 331 61 220.7 39.7 d-amphetamineOral d-amphetamine 1.5 142.2 0.5 446 461 84 307.3 94.8 Oral L-lysine- 3126.9 1.5 721 784 72 261.3 42.3 d-amphetamine Oral d-amphetamine 3 217.21.5 885 921 84 307.0 72.4 Oral L-lysine- 6 310.8 3 1,680 1,797 82 299.551.8 d-amphetamine Oral d-amphetamine 6 815.3 0.25 1,319 1,362 62 227.0135.9 Oral L-lysine- 12 412.6 5 2,426 2,701 62 225.1 34.4 d-amphetamineOral d-amphetamine 12 1,533.1 0.25 4,252 4,428 101 369.0 127.8 OralL-lysine- 60 2,164.3 5 9995.1 11.478 52 191.3 36.1 d-amphetamine Orald-amphetamine 60 13,735 1 32,323 48,707 223 811.8 228.9

TABLE 16 Pharmacokinetic Parameters of d-amphetamine Following BolusIntravenous Administration of L-lysine-d-amphetamine. Dose Cmax (mg/(ng/ Tmax AUC(0-24) AUC(inf) Route Drug kg) mL) (h) (ng · mL/h) (ng ·mL/h) IV L-lysine- 1.5 99.5 0.5 237.8 237.9 d-amphetamine IVd-amphetamine 1.5 420.2 0.083 546.7 546.9

TABLE 17 Pharmacokinetic Parameters of d-amphetamine FollowingIntranasal Administration of L-lysine-d-amphetamine. Dose (mg/ Cmax TmaxAUC(0-1) AUC(inf) Route Drug kg) (ng/mL) (h) (ng · mL/h) (ng · mL/h) INL-lysine-d- 10.16 78.6 1 56 91 amphetamine IN d- 4.12 1962.9 0.083 10327291 amphetamine

Tables 18-20 summarize the pharmacokinetic parameters ofL-lysine-d-amphetamine following oral, bolus intravenous, or intranasaladministration of L-lysine-d-amphetamine.

TABLE 18 Pharmacokinetic Parameters of L-lysine-d-amphetamine FollowingOral Administration of L-lysine-d-amphetamine at Escalating Doses. DoseCmax Tmax AUC (0-8) AUC (inf) Dose Drug (mg/kg) (ng/ml) (ng/ml) (ng ·ml/h) (ng · ml/h) F (%) Oral L-lysine-d-amphetamine 1.5 36.5 0.25 59.460 2.6 Oral L-lysine-d-amphetamine 3 135.4 1.5 329.7 332.1 7.2 OralL-lysine-d-amphetamine 6 676.8 0.25 1156.8 1170.8 12.8 OralL-lysine-d-amphetamine 12 855.9 1 4238.6 4510.4 24.6 OralL-lysine-d-amphetamine 60 1870.3 3 8234.3 8499.9 9.3

TABLE 19 Pharmacokinetic Parameters of L-lysine-d-amphetamine FollowingBolus Intravenous Administration of L-lysine-d-amphetamine. Dose (mg/Cmax Tmax AUC (0-24) AUC (inf) Route Drug kg) (ng/mL) (h) (ng · mL/h)(ng · mL/h) IV L-lysine- 1.5 4513.1 0.083 2,282 2,293 d- amphetamine

TABLE 20 Pharmacokinetic Parameters of L-lysine-d-amphetamine FollowingIntranasal Administration of L-lysine-d-amphetamine. Dose (mg/ Cmax TmaxAUC (0-1) AUC (inf) Route Drug kg) (ng/mL) (h) (ng · mL/h) (ng · mL/h)IN L-lysine- 3 3345.1 0.25 2,580 9,139 d- amphetamine

Tables 21 and 22 summarize the percent bioavailability of d-amphetaminefollowing oral, intranasal, or intravenous administration ofL-lysine-d-amphetamine as compared to d-amphetamine sulfate.

TABLE 21 Percent Bioavailability (AUC_(inf)) of d-amphetamine FollowingAdministration of L-lysine-d-amphetamine by Various Routes as Comparedto Bioavailability Following Administration of d-amphetamine Sulfate.Dose (mg/kg) d-amphetamine base 1.5 3 6 12 60 HED 19.9 39.9 79.7 159.4797.2 Oral 72 85 132 61 24 IV 43 NA NA NA NA IN NA 1 NA NA NA

TABLE 22 Percent Bioavailability (C_(max)) of d-amphetamine FollowingAdministration of L-lysine-d-amphetamine by Various Routes as Comparedto Bioavailability Following Administration of d-amphetamine Sulfate.Dose (mg/kg) d- amphetamine base 1.5 3 6 12 60 HED 19.9 39.9 79.7 159.4797.2 Oral 42 58 38 27 16 IV 24 NA NA NA NA IN NA 4 NA NA NA

Tables 23-28 summarize the time-course concentrations of d-amphetamineand L-lysine-d-amphetamine following oral, intranasal or intravenousadministration of either d-amphetamine or L-lysine-d-amphetamine.

TABLE 23 Time-course Concentrations of d-amphetamine Following BolusIntravenous Administration of L-lysine-d-amphetamine or d-amphetamineSulfate at Doses Containing 1.5 mg/kg d-amphetamine Base. Concentration(ng/ml) Time L-lysine- d-amphetamine (hours) d-amphetamine sulfate 0 0 00.083 52.8 420.2 0.5 99.5 249.5 1.5 47.1 97.9 3 21.0 38.3 5 9.0 13.2 83.7 4.3 24 0.1 0.2

TABLE 24 Time-course Concentrations of L-lysine-d-amphetamine FollowingBolus Intravenous Administration of L-lysine-d-amphetamine at a DoseContaining 1.5 mg/kg d-amphetamine Base. Concentration (ng/ml) TimeL-lysine- (hours) d-amphetamine 0 0 0.083 4513.1 0.5 1038.7 1.5 131.4 319.3 5 17.9 8 8.7 24 11.5

TABLE 25 Time-course Concentrations of d-amphetamine Following OralAdministration of L-lysine-d-amphetamine at Various Doses (mg/kgd-amphetamine base). Time Concentration (ng/ml) (hours) 1.5 mg/kg 3mg/kg 6 mg/kg 12 mg/kg 60 mg/kg 0 0 0 0 0 0 0.25 20.5 25.3 96 54.3 90.90.5 34 40.9 140.2 96 175.1 1 46.7 95.1 225.9 233.3 418.8 1.5 40.7 126.9268.4 266 440.7 3 59.6 105 310.8 356.8 1145.5 5 38.6 107.6 219.5 412.62164.3 8 17.1 48 86 225.1 1227.5

TABLE 26 Time-course Concentrations of d-amphetamine Following OralAdministration of d-amphetamine Sulfate at Various Doses (mg/kgd-amphetamine Base). Time Concentration (ng/ml) (hours) 1.5 mg/kg 3mg/kg 6 mg/kg 12 mg/kg 60 mg/kg 0 0 0 0 0 0 0.25 107.1 152.6 815.31533.1 6243.6 0.5 142.2 198.4 462.7 1216 7931.6 1 105.7 191.3 301.3828.8 13735.2 1.5 129.5 217.2 314 904.8 11514.9 3 52.6 135.3 134.6 519.9NA 5 29.5 73.5 77.4 404.3 NA 8 11.5 25.7 31.8 115.4 NA

TABLE 27 Time-course Concentrations of d-amphetamine FollowingIntranasal Administration of L-lysine-d-amphetamine or d-amphetamineSulfate at Doses Containing 3 mg/kg d-amphetamine Base. Concentration(ng/ml) Time L-lysine- d-amphetamine (hours) d-amphetamine sulfate 0 0 00.083 31.2 1962.9 0.25 45.3 1497.3 0.5 61.3 996.2 1 78.6 404.6 AUC 561032.3

TABLE 28 Time-course Concentrations of L-lysine-d-amphetamine FollowingIntranasal Administration of L-lysine-d-amphetamine at a Dose Containing3 mg/kg d-amphetamine Base. Conc. (ng/ml) L-lysine-d- Time (h)amphetamine 0 0 0.083 3345.1 0.25 3369.7 0.5 2985.8 1 1359.3

Example 19 LC/MS/MS Analysis of Bioavailability in Dogs ExampleExperimental Design:

This was a non-randomized, two-treatment crossover study. All animalswere maintained on their normal diet and were fasted overnight prior toeach dose administration. L-lysine-d-amphetamine dose was based on thebody weight measured on the morning of each dosing day. The actual dosedelivered was based on syringe weight before and after dosing. Serialblood samples were obtained from each animal by direct venipuncture of ajugular vein using vacutainer tubes containing sodium heparin as theanticoagulant. Derived plasma samples were stored frozen until shipmentto the Quest Pharmaceutical Services, Inc. (Newark, Del.).Pharmacokinetic analysis of the plasma assay results was conducted byCalvert. Animals were treated as follows:

Dose Dose # of Dog/ Route of Dose Conc. Vol. Level Sex AdministrationTreatment (mg/mL) (mL/kg) (mg/kg) 3M PO 1 0.2 10 2 3M IV 2 1 2 2The mg units in the dose concentration and dose level refer to the freebase form of test article.

Administration of the Test Article:

Oral: The test article was administered to each animal via a single oralgavage. On Day 1, animals received the oral dose by gavage using anesophageal tube attached to a syringe. Dosing tubes were flushed withapproximately 20 mL tap water to ensure the required dosing solution wasdelivered.

Intravenous: On Day 8, animals received L-lysine-d-amphetamine as asingle 30-minute intravenous infusion into a cephalic vein.

Sample Collection:

Dosing Formulations: Post-dosing, remaining dosing formulation was savedand stored frozen.

Blood: Serial blood samples (2 mL) were collected using venipuncturetubes containing sodium heparin. Blood samples were taken at 0, 0.25,0.5, 1, 2, 4, 8, 12, 24, 48, and 72 hours post-oral dosing. Bloodsamples were collected at 0, 0.167, 0.33, 0.49 (prior to stop ofinfusion), 0.583, 0.667, 0.75, 1, 2, 3, 4, 8, 12, and 23 hourspost-intravenous infusion start. Collected blood samples were chilledimmediately.

Plasma: Plasma samples were obtained by centrifugation of blood samples.Duplicate plasma samples (about 0.2 mL each) were transferred intoprelabeled plastic vials and stored frozen at approximately −70° C.

Sample Assay:

Plasma samples were analyzed for L-lysine-d-amphetamine andd-amphetamine using a validated LC-MS/MS method with an LLOQ of 1 ng/mLfor both analytes.

Microsoft Excel (Version 6, Microsoft Corp., Redmond, Wash.) was usedfor calculation of mean plasma concentration and graphing of the plasmaconcentration-time data. Pharmacokinetic analysis (non-compartmental)was performed using the WinNonlin® software program (Version 4.1,Pharsight, Inc. Mountain View, Calif.). The maximum concentration,C_(max), and the time to C_(max), T_(max), were observed values. Thearea under the plasma concentration-time curve (AUC) was determinedusing linear-log trapezoidal rules. The apparent terminal rate constant(λz) was derived using linear least-squares regression with visualinspection of the data to determine the appropriate number of points(minimum of 3 data points) for calculating λz. The AUC(0-inf) wascalculated as the sum of AUC(0-t) and Cpred/λz, where Cpred was thepredicted concentration at the time of the last quantifiableconcentration. The plasma clearance (CL/F) was determined as the ratioof Dose/AUC (0-inf). The mean residence time (MRT) was calculated as theratio of AUMC(0-inf)/AUC (0-inf), where AUMC(0-inf) was the area underthe first moment curve from the time zero to infinity. The volume ofdistribution at steady state (V_(ss)) was estimated as CL*MRT. Half-lifewas calculated as ln 2/λz. The oral bioavailability (F) was calculatedas the ratio of AUC(0-inf) following oral dosing to AUC(0-inf) followingintravenous dosing. Descriptive statistics (mean and standard deviation)of the pharmacokinetic parameters were calculated using Microsoft Excel.

The objectives of this study were to characterize the pharmacokineticsof L-lysine-d-amphetamine and d-amphetamine following administration ofL-lysine-d-amphetamine in male beagle dogs. As shown in FIG. 27, in across-over design, L-lysine-d-amphetamine was administered to 3 malebeagle dogs orally (2 mg/kg) and intravenously (2 mg/kg, 30-minuteinfusion). Blood samples were collected up to 24 and 72 hour after theintravenous and oral does, respectively. Plasma samples were analyzedusing a LC-MS/MS assay which provided an LLOQ of 1 ng/mL for bothanalytes.

The mean L-lysine-d-amphetamine and d-amphetamine plasmaconcentration-time profiles following an intravenous or oral dose ofL-lysine-d-amphetamine are presented in FIGS. 29 and 30, respectively.Comparative profiles of L-lysine-d-amphetamine to d-amphetaminefollowing both routes are depicted in FIGS. 27-28. Individual plots aredepicted in FIGS. 31-32. The pharmacokinetic parameters are summarizedin Tables 29-37.

Following a 30-minute intravenous infusion of L-lysine-d-amphetamine,the plasma concentration reached a peak at the end of the infusion.Post-infusion L-lysine-d-amphetamine concentration declined very rapidlyin a biexponential manner, and fell below the quantifiable limit (1ng/mL) by approximately 8 hours post-dose. Results of non-compartmentalpharmacokinetic analysis indicate that L-lysine-d-amphetamine is a highclearance compound with a moderate volume of distribution (Vss)approximating total body water (0.7 L/kg). The mean clearance value was2087 mL/h·kg (34.8 mL/min·kg) and was similar to the hepatic blood flowin the dog (40 mL/min·kg). Consequently, L-lysine-d-amphetamine is amoderate to high hepatic extraction compound with significant first passeffects (including the conversion to d-amphetamine) following oraladministration.

L-lysine-d-amphetamine was rapidly absorbed after oral administrationwith T_(max) at 0.5 hours in all three dogs. Mean absolute oralbioavailability was 33%. Since significant first pass effects areexpected for L-lysine-d-amphetamine, a 33% bioavailability suggests thatL-lysine-d-amphetamine is very well absorbed in the dog. The apparentterminal half-life was 0.39 hours, indicating rapid elimination, asobserved following intravenous administration.

Plasma concentration-time profiles of d-amphetamine followingintravenous or oral administration of L-lysine-d-amphetamine were verysimilar, with C_(max), T_(max) and AUC values for both routesessentially the same. At a 2 mg/kg oral dose of L-lysine-d-amphetamine,the mean C_(max) of d-amphetamine was 104.3 ng/mL. The half-life ofd-amphetamine was 3.1 to 3.5 hours, much longer when compared toL-lysine-d-amphetamine.

In this study, L-lysine-d-amphetamine was infused over a 30 minute timeperiod. Due to rapid clearance of L-lysine-d-amphetamine it is likelythat bioavailability of d-amphetamine from L-lysine-d-amphetamine woulddecrease if a similar dose were given by intravenous bolus injection.Even when given as an infusion the bioavailability of d-amphetamine fromL-lysine-d-amphetamine did not exceed that of a similar dose givenorally and the time to peak concentration was substantially delayed.This data further supports that L-lysine-d-amphetamine affords adecrease in the abuse liability of d-amphetamine by intravenousinjection.

TABLE 29 Pharmacokinetic Parameters of L-lysine-d-amphetamine in MaleBeagle Dogs Following Oral or Intravenous Administration ofL-lysine-d-amphetamine (1 mg/kg d-amphetamine base). Dose C_(max)T_(max) ^(a) AUC (inf) t_(1/2) MRT CL/F V_(ss) F Route (mg/kg) (ng/mL)(h) (ng · h/mL) (h) (h) (mL/h · kg) (mL/kg) (%) IV 1 1650  0.49 964 0.880.33 2087  689 NA (0.00) (178) (0.49-0.49) (97.1) (0.2) (0.03) (199)(105.9) Oral 1   328.2 0.5  319 0.39 0.81 6351  NA 33 (0.00)   (91.9)(0.5-0.5) (46.3) (0.1) (0.19)   (898.3) (1.9) ^(a)median (range)

TABLE 30 Pharmacokinetic Parameters of d-amphetamine in Male Beagle DogsFollowing Oral or Intravenous Administration of L-lysine-d-amphetamine(1 mg/kg d-amphetamine base). Dose C_(max) T_(max) ^(a) AUC (inf)t_(1/2) Route (mg/kg) (ng/mL) (h) (ng · h/mL) (h) IV 2 113.2 1.0 672.53.14 (0.00) (3.2) (0.67-2.0)  (85.7) (0.4) Oral 2 104.3 2.0 728.0 3.48(0.00) (21.8) (2-2) (204.9) (0.4) ^(a)median (range)

TABLE 31 Pharmacokinetics of L-lysine-d-amphetamine in Male Beagle DogsFollowing Intravenous Administration of L-lysine-d-amphetamine (1 mg/kgd-amphetamine base). Dose Route: 30-min iv Infusion Dose: 2 mg/kg/h(free form) C_(max) T_(max) ^(a) AUC (0-t) AUC (inf) t_(1/2) CL V_(ss)MRT Dog ID (ng/mL) (h) (ng · h/mL) (ng · h/mL) (h) (mL/h/kg) (mL/kg) (h)1 1470.3 0.49 898.2 900.2 0.72 2222 807.4 0.36 2 1826.4 0.49 1072.31076.1 ND^(b) 1859 603.4 0.32 3 1654.2 0.49 914.1 916.9 1.05 2181 656.00.30 Mean 1650 0.49 961.5 964.4 0.88 2087 689.0 0.33 SD 178 0.49-0.4996.0 97.1 0.2 199 105.9 0.03 ^(a)median (range); ^(b)not determinedAbbreviations of pharmacokinetic parameters are as follows: C_(max),maximum observed plasma concentration; AUC (0-t), total area under theplasma concentration versus time curve from 0 to the last data point;AUC (0-inf), total area under the plasma concentration versus timecurve; t_(1/2), apparent terminal half-life; CL, clearance following ivadministration; MRT, mean residence time; V_(ss), volume of distributionat steady state.

TABLE 32 Pharmacokinetic Parameters of L-lysine-d-amphetamine in MaleBeagle Dogs Following Oral Administration of L-lysine-d-amphetamine (1mg/kg d-amphetamine base). Dose Route: Oral Dose: 2 mg/kg (free form)C_(max) T_(max) ^(a) AUC (0-t) AUC (inf) t_(1/2) CL/F MRT F Dog ID(ng/mL) (h) (ng · h/mL) (ng · h/mL) (h) (mL/h/kg) (h) (%) 1 350.2 0.5275.3 277.1 0.24 7218 0.68 30.8 2 407.2 0.5 367.8 368.7 0.48 5424 0.7434.3 3 227.4 0.5 310.8 312.0 0.45 6410 1.03 34.0 Mean 328.2 0.5 318.0319.3 0.39 6351 0.81 33.0 SD 91.9 0.0 46.7 46.3 0.1 898.3 0.19 1.9^(a)median (range) Abbreviations of pharmacokinetic parameters are asfollows: C_(max), maximum observed plasma concentration; T_(max), timewhen C_(max) observed; AUC (0-t), total area under the plasmaconcentration versus time curve from 0 to the last data point; AUC(0-inf), total area under the plasma concentration versus time curve;t_(1/2), apparent terminal half life; CL/F, oral clearance; MRT, meanresidence time; F, bioavailability.

TABLE 33 Pharmacokinetics of L-lysine-d-amphetamine in Male Beagle DogsFollowing Intravenous Administration of L-lysine-d-amphetamine (1 mg/kgd-amphetamine base). Dose Route: 30-min iv Infusion Dose: 2 mg/kg ofL-lysine-d-amphetamine (free form) C_(max) T_(max) ^(a) AUC (0-t) AUC(inf) t_(1/2) Dog ID (ng/mL) (h) (ng · h/mL) (ng · h/mL) (h) 1 111.2 2.0751.9 757.6 3.35 2 116.8 0.67 668.5 673.7 3.43 3 111.4 1.0 557.8 586.12.65 Mean 113.2 1.00 659.4 672.5 3.14 SD 3.2 0.67-2.0 97 85.7 0.4^(a)median (range) Abbreviations of pharmacokinetic parameters are asfollows: C_(max), maximum observed plasma concentration; T_(max), timewhen C_(max) observed; AUC (0-t), total area under the plasmaconcentration versus time curve from 0 to the last data point; AUC(0-inf), total area under the plasma concentration versus time curve;t_(1/2), apparent terminal half-life; CL/F, oral clearance; MRT, meanresidence time; F, bioavailability.

TABLE 34 Pharmacokinetics of L-lysine-d-amphetamine in Male Beagle DogsFollowing Oral Administration of L-lysine-d-amphetamine (1 mg/kgd-amphetamine base). Dose Route: Oral Dose: 2 mg/kg ofL-lysine-d-amphetamine (free form) C_(max) T_(max) ^(a) AUC (0-t) AUC(inf) t_(1/2) Dog ID (ng/mL) (h) (ng · h/mL) (ng · h/mL) (h) 1 102.1 2.0686.34 696.89 3.93 2 127.2 2.0 937.57 946.62 3.44 3 83.7 2.0 494.61540.38 3.06 Mean 104.3 2.0 706.2 728.0 3.48 SD 21.8 2.0-2.0 222.1 204.90.4 ^(a)median (range) Abbreviations of pharmacokinetic parameters areas follows: C_(max), maximum observed plasma concentration; T_(max),time when C_(max) observed; AUC (0-t), total area under the plasmaconcentration versus time curve from 0 to the last data point; AUC(0-inf), total area under the plasma concentration versus time curve;t_(1/2), apparent terminal half-life; CL/F, oral clearance; MRT, meanresidence time; F, bioavailability.

TABLE 35 Pharmacokinetics of d-amphetamine in Male Beagle Dogs FollowingOral Administration of L-lysine-d-amphetamine or d-amphetamine sulfate(1.8 mg/kg d-amphetamine base). Mean Plasma Concentration StandardDeviation (SD) Coefficient of Variation (CV) Time L-lysine-d-L-lysine-d- L-lysine-d (hours) d-amphetamine amphetamine d-amphetamineamphetamine d-amphetamine amphetamine 0 0 0 0 0 0 0 1 431.4 223.7 140.795.9 32.6 42.9 2 360 291.8 87.6 93.6 24.3 32.1 4 277.7 247.5 68.1 6624.5 26.7 6 224.1 214.7 59.3 62.1 26.5 28.9 8 175.4 150 66.7 40.1 38.026.7 12 81.4 47.6 58.7 19 72.1 39.9 16 33 19.6 28.1 9 85.2 45.9 24 7.24.5 4.5 1.7 62.5 37.8

TABLE 36 Pharmacokinetics of d-amphetamine in Female Beagle DogsFollowing Oral Administration of L-lysine-d-amphetamine or d-amphetaminesulfate (1.8 mg/kg d- amphetamine base). Mean Plasma Standard DeviationCoefficient of Concentration (SD) Variation (CV) Time L-lysine-d-L-lysine-d- L-lysine-d- (hours) d-amphetamine amphetamine d-amphetamineamphetamine d-amphetamine amphetamine 0 0 0 0 0 0 0 1 217.8 308.8 141.740.7 65.1 13.2 2 273.5 308 113.7 29.6 41.6 9.6 4 266 260.9 132.7 37.349.9 14.3 6 204.7 212.1 84.5 38.7 41.3 18.2 8 160.1 164.3 72.7 43.5 45.426.5 12 79.4 68.7 41.3 31 52.0 45.1 16 25.5 22.3 13.4 4.7 52.5 21.1 245.6 5.4 4.1 1.9 73.2 35.2

TABLE 37 Pharmacokinetic Parameters of d-amphetamine in Male and FemaleBeagle Dogs Following Oral Administration of L-lysine-d-amphetamine ord-amphetamine sulfate (1.8 mg/kg d-amphetamine base). Males FemalesCompound Compound d- L-lysine-d- d- L-lysine-d- Parameter amphetamineamphetamine amphetamine amphetamine AUCinf 3088.9 2382.2 2664.5 2569.9Percent 100 77 100 96 Cmax 431.4 291.8 308.8 273.5 Percent 100 67 100 89Tmax (hours) 1 2 1 2 Percent 100 200 100 200

Example 20 Delayed Cardiovascular Effects of L-lysine-d-amphetamine asCompared to d-amphetamine Following Intravenous Infusion

Systolic and diastolic blood pressure (BP) are increased byd-amphetamine even at therapeutic doses. Since L-lysine-d-amphetamine isexpected to release d-amphetamine (albeit slowly) as a result ofsystemic metabolism, a preliminary study was done using equimolar dosesof d-amphetamine or L-lysine-d-amphetamine to 4 dogs (2 male and 2female). The results suggest that the amide prodrug is inactive and thatslow release of some d-amphetamine, occurs beginning 20 minutes afterthe first dose. Relative to d-amphetamine, however, the effects are lessrobust. For example, the mean blood pressure is graphed in FIG. 35.Consistent with previously published data (Kohli and Goldberg, 1982),small doses of d-amphetamine were observed to have rapid effects onblood pressure. The lowest dose (0.202 mg/kg, equimolar to 0.5 mg/kg ofL-lysine-d-amphetamine) produced an acute doubling of the mean BPfollowed by a slow recovery over 30 minutes.

By contrast, L-lysine-d-amphetamine produced very little change in meanBP until approximately 30 minutes after injection. At that time,pressure increased by about 20-50%. Continuous release of d-amphetamineis probably responsible for the slow and steady increase in bloodpressure over the remaining course of the experiment. Upon subsequentinjections, d-amphetamine is seen to repeat its effect in a non-dosedependent fashion. That is, increasing dose 10-fold from the firstinjection produced a rise to the same maximum pressure. This may reflectthe state of catecholamine levels in nerve terminals upon successivestimulation of d-amphetamine bolus injections. Note that the rise inmean blood pressure seen after successive doses ofL-lysine-d-amphetamine (FIG. 35) produces a more gradual and lessintense effect. Similar results were observed for left ventricularpressure (FIG. 36). These results further substantiate the significantdecrease in d-amphetamine bioavailability by the intravenous route whengiven as L-lysine-d-amphetamine. As a result the rapid onset of thepharmacological effect of d-amphetamine that is sought by personsinjecting the drug is eliminated.

TABLE 38 Effects of L-lysine-d-amphetamine on Cardiovascular Parametersin the Anesthetized Dog - Mean Values (n = 2) % % % % TREATMENT TIME SAPChange DAP Change MAP Change LVP Change 0.9% Saline 0 81 0 48 0 61 0 870 1 ml/kg 30 87 7 54 11 67 10 87 0 L-lysine-d- 0 84 0 51 0 64 0 86 0amphetamine 0.5 mg/kg 5 87 4 52 3 66 3 87 2 15 93 11 51 1 67 5 95 11 25104 25 55 8 73 15 105 22 30 107 28 58 14 77 21 108 26 L-lysine-d- 0 1050 55 0 74 0 108 0 amphetamine 1.0 mg/kg 5 121 15 63 15 85 15 120 11 15142 35 73 33 100 35 140 29 25 163 55 97 75 124 68 162 50 30 134 28 73 3298 32 144 33 L-lysine-d- 0 132 0 71 0 95 0 144 0 amphetamine 5.0 mg/kg 5142 7 71 0 99 4 151 5 15 176 33 98 39 130 37 184 28 25 126 −5 69 −3 96 1160 11 30 132 0 70 −1 99 4 163 13 SAP—systolic arterial pressure (mmHg)MAP—mean arterial pressure (mmHg) DAP—diastolic arterial pressure (mmHg)LVP—left ventricular pressure (mmHg) % Change—percent change fromrespective Time 0.

TABLE 39 Effects of d-Amphetamine on Cardiovascular Parameters in theAnesthetized Dog - Mean Values (n = 2) % % % % TREATMENT TIME SAP ChangeDAP Change MAP Change LVP Change 0.9% Saline 0 110 0 67 0 84 0 105 0 1ml/kg 30 108 −2 65 −3 82 −2 101 −3 d-amphetamine 0 111 0 67 0 84 0 104 00.202 mg/kg 5 218 97 145 117 176 109 214 107 15 168 52 97 45 125 49 15752 25 148 34 87 30 110 31 142 37 30 140 26 80 20 103 23 135 30d-amphetamine 0 139 0 78 0 101 0 133 0 0.404 mg/kg 5 240 73 147 88 18785 238 79 15 193 39 112 44 145 43 191 43 25 166 19 92 17 122 20 168 2630 160 16 87 11 117 16 163 22 d-amphetamine 0 158 0 87 0 115 0 162 02.02 mg/kg 5 228 44 128 48 169 47 227 40 15 196 24 107 23 142 23 200 2425 189 20 102 17 135 17 192 19 30 183 16 98 13 129 12 187 16SAP—systolic arterial pressure (mmHg) MAP—mean arterial pressure (mmHg)DAP—diastolic arterial pressure (mmHg) LVP—left ventricular pressure(mmHg) % Change—percent change from respective Time 0.

Example 21 Pharmacodynamic (Locomotor) Response to Amphetamine vs.L-lysine-d-amphetamine by Oral Administration

Male Sprague-Dawley rats were provided water ad libitum, fastedovernight and dosed by oral gavage with 6 mg/kg of amphetamine orL-lysine-d-amphetamine containing the equivalent amount ofd-amphetamine. Horizontal locomotor activity (HLA) was recorded duringthe light cycle using photocell activity chambers (San DiegoInstruments). Total counts were recorded every 12 minutes for theduration of the test. Rats were monitored in three separate experimentsfor 5, 8, and 12 hours, respectively. Time vs. HLA counts ford-amphetamine vs. L-lysine-d-amphetamine is shown in FIGS. 37-38. Ineach experiment the time until peak activity was delayed and thepharmacodynamic effect was evident for an extended period of time forL-lysine-d-amphetamine as compared to d-amphetamine. The total activitycounts for HLA of Lys-Amp dosed rats were increased (11-41%) over thoseinduced by d-amphetamine in all three experiments (Tables 40 and 41).

TABLE 40 Locomotor Activity of Rats Orally Administered d-amphetaminevs. L-lysine- d-amphetamine (5 Hours) Total Activity Peak of activityTime of Peak Time of Last Total Activity Counts Above (Counts per(Counts per Count Above 200 Test Material Counts Baseline 0.2 h) 0.2 h)per 0.2 h Vehicle 4689 4174 80 1.4 — L-lysine-d- 6417 5902 318 1.8   5 hamphetamine d-amphetamine 515 0 291 0.6 2.6 h

TABLE 41 Locomotor Activity of Rats Orally Administered Amphetamine vs.L-lysine-d- amphetamine (12 Hours) Total Activity Peak of activity Timeof Peak Time of Last Total Activity Counts Above (Counts per (Counts perCount Above 100 Test Material Counts Baseline 0.2 h) 0.2 h) per 0.2 hVehicle 936 0 81 7.2 — L-lysine-d- 8423 7487 256 1.8 8.6 h amphetamined-amphetamine 6622 5686 223 0.6 6.4 h

Example 22 Pharmacodynamic Response to Amphetamine vs.L-lysine-d-amphetamine by Intranasal Administration

Male Sprague-Dawley rats were dosed by intranasal administration with1.0 mg/kg of amphetamine or L-lysine-d-amphetamine containing theequivalent amount of d-amphetamine. In a second set of similarly dosedanimals carboxymethyl cellulose (CMC) was added to the drug solutions ata concentration of 62.6 mg/ml (approximately 2-fold higher than theconcentration of L-lysine-d-amphetamine and 5-fold higher than thed-amphetamine content). The CMC drug mixtures were suspended thoroughlybefore each dose was delivered. Locomotor activity was monitored usingthe procedure described in the section titled example 7. As shown inFIGS. 39-40, the activity vs. time (1 hour or 2 hours) is shown foramphetamine/CMC vs. L-lysine-d-amphetamine and compared to that ofamphetamine vs. L-lysine-d-amphetamine CMC. As seen in FIG. 39, additionof CMC to L-lysine-d-amphetamine decreased the activity response of INdosed rats to levels similar to the water/CMC control, whereas no effectwas seen on amphetamine activity by the addition of CMC. The increase inactivity over baseline of L-lysine-d-amphetamine with CMC was only 9%compared to 34% for Lys-Amp without CMC when compared to activityobserved for d-amphetamine dosed animals (Table 42). CMC had noobservable affect on d-amphetamine activity induced by INadministration.

TABLE 42 Locomotor Activity of Intranasal d-amphetamine vs.L-lysine-d-amphetamine with and without CMC Total Activity TotalActivity Counts Counts Percent Drug n (1 h) Above Baseline d-amphetamined-mphetamine 3 858 686 100 d-amphetamine CMC 3 829 657 100 L-lysine-d- 4408 237 35 amphetamine L-lysine-d- 4 232 60 9 amphetamine CMC Water 1172 0 0 Water CMC 1 172 0 0

Example 23 Pharmacodynamic Response to Amphetamine vs.L-lysine-d-amphetamine by Intravenous (IV) Administration

Male Sprague-Dawley rats were dosed by intravenous administration with1.0 mg/kg of d-amphetamine or L-lysine-d-amphetamine containing theequivalent amount of amphetamine. The activity vs. time (3 hours) isshown for d-amphetamine vs. L-lysine-d-amphetamine (FIG. 41). Theactivity induced by L-lysine-d-amphetamine was substantially decreasedand time to peak activity was delayed. The activity expressed as totalactivity counts over a three hour period of time is shown in FIG. 41.The increase in activity over baseline of L-lysine-d-amphetamine was 34%for L-lysine-d-amphetamine when compared to activity observed ford-amphetamine dosed animals (Table 43).

TABLE 43 Total activity counts after d-amphetamine vs.L-lysine-d-amphetamine Following Intravenous (IV) Administration. TotalActivity Counts Above Percent Drug n 3 h Baseline d-amphetamined-amphetamine 3 1659 1355 100 L-lysine-d- 4 767 463 34 amphetamine Water1 304 0 0

Example 24 Decrease in Toxicity of Orally AdministeredL-lysine-d-amphetamine

Three male and three female Sprague Dawley rats per group were given asingle oral administration of L-lysine-d-amphetamine at 0.1, 1.0, 10,60, 100 or 1000 mg/kg (Table 44). Each animal was observed for signs oftoxicity and death on Days 1-7 (with Day 1 being the day of the dose)and one rat/sex/group was necropsied upon death (scheduled orunscheduled).

TABLE 44 Dosing Chart Oral Administration of L-lysine-d-amphetamineToxicity Testing. No. of Concen- Animals Dosages trations Groups M FTest Article (mg/kg) (mg/mL) 1 3 3 L-lysine-d-amphetamine 0.1 0.01 2 3 3L-lysine-d-amphetamine 1.0 0.1 3 3 3 L-lysine-d-amphetamine 10 1.0 4 3 3L-lysine-d-amphetamine 60 6.0 5 3 3 L-lysine-d-amphetamine 100 10 6 3 3L-lysine-d-amphetamine 1000 100

Key observations of this study include:

-   -   All animals in Groups 1-3 showed no observable signs throughout        the conduct of the study.    -   All animals in Groups 4-6 exhibited increased motor activity        within two hours post-dose and which lasted into Day 2.    -   One female rat dosed at 1000 mg/kg was found dead on Day 2.        Necropsy revealed chromodacryorrhea, chromorhinorrhea, distended        stomach (gas), enlarged adrenal glands, and edematous and        distended intestines.    -   A total of 4 rats had skin lesions of varying degrees of        severity on Day 3.    -   One male rat dosed at 1000 mg/kg was euthanatized on Day 3 due        to open skin lesions on the ventral neck.    -   All remaining animals appeared normal from Day 4 through Day 7.

Animals were observed for signs of toxicity at 1, 2 and 4 h post-dose,and once daily for 7 days after dosing and cage-side observations wererecorded. Animals found dead, or sacrificed moribund were necropsied anddiscarded. A total of one animal/sex/group was necropsied upon scheduledor unscheduled death.

Cage-side observations and gross necropsy findings are summarized inTable 5. The data are not sufficient to establish a lethal dose,however, the study indicates that the lethal oral dose ofL-lysine-d-amphetamine is above 1000 mg/kg, because only one deathoccurred out of a group of six animals. Although a second animal in thisdose group was euthanatized on Day 3, it was done for humane reasons andit was felt that this animal would have fully recovered. Observationssuggested drug-induced stress in Groups 4-6 that is characteristic ofamphetamine toxicity (NTP, 1990; NIOSH REGISTRY NUMBER: SI1750000;Goodman et. al., 1985). All animals showed no abnormal signs on Days 4-7suggesting full recovery at each treatment level.

The lack of data to support an established lethal dose is believed to bedue to a putative protective effect of conjugating amphetamine withlysine. Intact L-lysine-d-amphetamine has been shown to be inactive, butbecomes active upon metabolism into the unconjugated form(d-amphetamine). Thus, at high doses, saturation of metabolism ofL-lysine-d-amphetamine into the unconjugated form may explain the lackof observed toxicity, which was expected at doses greater than 100mg/kg, which is consistent with d-amphetamine sulfate (NTP, 1990). Theformation rate of d-amphetamine and the extent of the formation ofamphetamine may both attribute to the reduced toxicity. Alternatively,oral absorption of L-lysine-d-amphetamine may also be saturated at suchhigh concentrations, which may suggest low toxicity due to limitedbioavailability of L-lysine-d-amphetamine.

Example 25 In Vitro Assessment of L-lysine-d-amphetamine PharmacodynamicActivity

It was anticipated that the acylation of amphetamine, as in the aminoacid conjugates discussed here, would significantly reduce the stimulantactivity of the parent drug. For example, Marvola (1976) showed thatN-acetylation of amphetamine completely abolished the locomotor activityincreasing effects in mice. To confirm that the conjugate was notdirectly acting as a stimulant, we tested (Novascreen, Hanover, Md.) thespecific binding of Lys-Amp (10⁻⁹ to 10⁻⁵ M) to human recombinantdopamine and norepinephrine transport binding sites using standardradioligand binding assays. The results (see Table 45) indicate that theLys-Amp did not bind to these sites. It seems unlikely that theconjugate retains stimulant activity in light of these results.(Marvola, M. (1976). “Effect of acetylated derivatives of somesympathomimetic amines on the acute toxicity, locomotor activity andbarbiturate anesthesia time in mice.” Acta Pharmacol Toxicol (Copenh)38(5): 474-89).

TABLE 45 Results From Radioligand Binding Experiments withL-lysine-d-amphetamine Reference Ki (M) for Assay Radioligand CompoundRef. Cpd. Activity* NE Transporter [3H]-Nisoxetine Desipramine 4.1 ×10⁻⁹ No DA Transporter [3H]-WIN35428 GBR-12909 7.7 × 10⁻⁹ No *Noactivity is defined as producing between −20% and 20% inhibition ofradioligand binding (Novascreen).

Example 26 In Vitro Assessment “Kitchen Tests” to Release Amphetamine

It was anticipated that attempts would be made by illicit chemists totreat the compound with various easily accessible physical and chemicalmethods by which to release free amphetamine from the conjugate. Anabuse-resistant preparation would have the additional feature of notreleasing d-amphetamine when exposed to water, acid (vinegar), base(baking powder and baking soda), and heat. In several tests withL-lysine-d-amphetamine and GGG-Amp, no amphetamine was detected afterthe following treatments:

Vinegar Tap Water Baking Powder Baking Soda L-lysine-d- 0% 0% 0% 0%amphetamine Gly₃-Amp 0% 0% 0% 0%Samples were heated to boiling for 20-60 minutes in each test.

Example 27 Bioavailability of Various Amino Acid-Amphetamine CompoundsAdministered by Oral, Intranasal, and Intravenous Routes

Oral Administration. Male Sprague-Dawley rats were provided water adlibitum, fasted overnight, and dosed by oral gavage with amphetamine oramino acid-amphetamine conjugates containing the equivalent amount ofamphetamine.

Intranasal Administration. Male Sprague-Dawley rats were dosed byintranasal administration with 1.8 mg/kg of amphetamine orlysine-amphetamine containing the equivalent amount of amphetamine.

The relative in vivo performance of various amino acid-amphetaminecompounds is shown in FIGS. 42-50 and summarized in Table 46. Intranasalbioavailability of amphetamine from Ser-Amp was decreased to some degreerelative to free amphetamine. However, this compound was notbioequivalent with amphetamine by the oral route of administration.Phenylalanine was bioequivalent with amphetamine by the oral route ofadministration, however, little or no decrease in bioavailability byparenteral routes of administration was observed. Gly₃-Amp had nearlyequal bioavailability (90%) by the oral route accompanied by a decreasein Cmax (74%). Additionally, Gly₃-Amp showed a decrease inbioavailability relative to amphetamine by intranasal and intravenousroutes.

TABLE 46 Percent Bioavailability of Amino Acid Amphetamine CompoundsAdministered by Oral, Intranasal or Intravenous Routes Oral IntranasalIntravenous Drug Percent AUC Percent Cmax Percent AUC Percent CmaxPercent AUC Percent Cmax Amphetamine 100 100 100 100 100 100 E-Amp 73 95NA NA NA NA EE-Amp 26 74 NA NA NA NA L-Amp 65 81 NA NA NA NA S-Amp 79/55 62/75 76 65 NA NA GG-Amp 79 88 88 85 NA NA GGG-Amp 111/68 74/7332 38 45 46 F-Amp 95 91 97 95 87 89 EEF-Amp 42 73 39 29 NA NA FF-Amp 2764 NA NA NA NA Gulonate-Amp 1 1 0.4 0.5 3 5 K-Amp 98 55 0.5 0.5 3 3KG-Amp 69 71 13 12 NA NA dK/K-Amp 16 7 2 2 NA NA LE-Amp 40 28 6 6 NA NAH-Amp 16 21 22 42 NA NA

Example 28 Decreased Oral C_(max) of d-Amphetamine Conjugates

Male Sprague-Dawley rats were provided water ad libitum, fastedovernight and dosed by oral gavage with amphetamine conjugate ord-amphetamine sulfate. All doses contained equivalent amounts ofd-amphetamine base. Plasma d-amphetamine concentrations were measured byELISA (Amphetamine Ultra, 109319, Neogen, Corporation, Lexington, Ky.).The assay is specific for d-amphetamine with only minimal reactivity(0.6%) of the major d-amphetamine metabolite(para-hydroxy-d-amphetamine) occurring. Plasma d-amphetamine andL-lysine-d-amphetamine concentrations were measured by LC/MS/MS whereindicated in examples.

Example 29 Decreased Intranasal Bioavailability (AUC and C_(max)) ofd-Amphetamine Conjugates

Male Sprague-Dawley rats were provided water ad libitum and doses wereadministered by placing 0.02 ml of water containing amphetamineconjugate or d-amphetamine sulfate into the nasal flares. All dosescontained equivalent amounts of d-amphetamine base. Plasma d-amphetamineconcentrations were measured by ELISA (Amphetamine Ultra, 109319,Neogen, Corporation, Lexington, Ky.). The assay is specific ford-amphetamine with only minimal reactivity (0.6%) of the majord-amphetamine metabolite (para-hydroxy-d-amphetamine) occurring. Plasmad-amphetamine and L-lysine-d-amphetamine concentrations were measured byLC/MS/MS where indicated in examples.

Example 30 Decreased Intravenous Bioavailability (AUC and C_(max)) ofd-Amphetamine Conjugates

Male Sprague-Dawley rats were provided water ad libitum and doses wereadministered by intravenous tail vein injection of 0.1 ml of watercontaining amphetamine conjugate or d-amphetamine sulfate. All dosescontained equivalent amounts of d-amphetamine base. Plasma d-amphetamineconcentrations were measured by ELISA (Amphetamine Ultra, 109319,Neogen, Corporation, Lexington, Ky.). The assay is specific ford-amphetamine with only minimal reactivity (0.6%) of the majord-amphetamine metabolite (para-hydroxy-d-amphetamine) occurring. Plasmad-amphetamine and L-lysine-d-amphetamine concentrations were measured byLC/MS/MS where indicated in examples.

Example 31 Attachment of Amphetamine to Variety of Chemical Moieties

The above examples demonstrate the use of an amphetamine conjugated to achemical moiety, such as an amino acid, which is useful in reducing thepotential for overdose while maintaining its therapeutic value. Theeffectiveness of binding amphetamine to a chemical moiety wasdemonstrated through the attachment of amphetamine to lysine (K),however, the above examples are meant to be illustrative only. Theattachment of amphetamine to any variety of chemical moieties (i.e.peptides, glycopeptides, carbohydrates, nucleosides, or vitamins)asdescribed below through similar procedures using the following exemplarystarting materials.

Amphetamine Synthetic Examples Synthesis of Gly₂-Amp

-   -   Gly₂-Amp was synthesized by a similar method except the amino        acid starting material was Boc-Gly-Gly-OSu.

Synthesis of Glu₂-Phe-Amp

-   -   Glu₂-Phe-Amp was synthesized by a similar method except the        amino acid starting material was Boc-Glu(OtBu)-Glu(OtBu)-OSu and        the starting drug conjugate was Phe-Amp (see Phe-Amp synthesis).

Synthesis of His-Amp

-   -   His-Amp was synthesized by a similar method except the amino        acid starting material was Boc-His(Trt)-OSu.

Synthesis of Lys-Gly-Amp

-   -   Lys-Gly-Amp was synthesized by a similar method except the amino        acid starting material was Boc-Lys(Boc)-OSu and the starting        drug conjugate was Gly-Amp (see Gly-Amp synthesis).

Synthesis of Lys-Glu-Amp

-   -   Lys-Glu-Amp was synthesized by a similar method except the amino        acid starting material was Boc-Lys(Boc)-OSu and the starting        drug conjugate was Glu-Amp.

Synthesis of Glu-Amp

-   -   Glu-Amp was synthesized by a similar method except the amino        acid starting material was Boc-Glu(OtBu)-OSu.        Synthesis of (d)-Lys-(l)-Lys-Amp    -   (d)-Lys-(l)-Lys-Amp was synthesized by a similar method except        the amino acid starting material was        Boc-(d)-Lys(Boc)-(l)-Lys(Boc)-OSu.

Synthesis of Gulonic Acid-Amp

-   -   Gul-Amp was synthesized by a similar method except the        carbohydrate starting material was gulonic acid-OSu.

Example 32 Lack of Detection of L-lysine-d-amphetamine in Brain TissueFollowing Oral Administration

Male Sprague-Dawley rats were provided water ad libitum, fastedovernight and dosed by oral gavage with L-lysine-d-amphetamine ord-amphetamine sulfate. All doses contained equivalent amounts ofd-amphetamine base. As shown in FIGS. 51A-B, similar levels ofd-amphetamine were detected in serum as well as in brain tissuefollowing administration of d-amphetamine sulfate orL-lysine-d-amphetamine. The conjugate L-lysine-d-amphetamine, however,was present in appreciable amounts in serum but was not detected inbrain tissue indicating that the conjugate does not cross the bloodbrain barrier to access the central nervous system site of action.

Example 33 Clinical Pharmacokinetic Evaluation and Oral Bioavailabilityof L-lysine-d-amphetamine Compared to Amphetamine Extended ReleaseProducts Adderall XR® and Dexadrine Spansule® Used in the Treatment ofADHD

TABLE 47 Treatment Groups and Dosage for Clinical PharmacokineticEvaluation of L-lysine-d-amphetamine Compared to Adderall XR ® orDexadrine Spansule ® Number Dose Treatment of Dose (amphetamine DrugGroup Subjects Dose (mg) base) L-lysine- A 10 1 × 25 mg 25 7.37d-amphetamine capsule L-lysine- B 10 3 × 25 mg 75 22.1 d-amphetaminecapsules Dexedrine C 10 3 × 10 mg 30 22.1 Spansule ® capsules AdderallXR ® D 10 1 × 30 mg 35 21.9 capsules plus 1 × 5 mg capsule

A clinical evaluation of the pharmacokinetics and oral bioavailabilityof L-lysine-d-amphetamine in humans was conducted.L-lysine-d-amphetamine was orally administered at doses approximatingthe lower (25 mg) and higher (75 mg) end of the therapeutic range basedon d-amphetamine base content of the doses. Additionally, the higherdose was compared to doses of Adderall XR® (Shire) or DexadrineSpansule® (GlaxoSmithKline) containing equivalent amphetamine base tothat of the higher L-lysine-d-amphetamine dose. Treatment groups anddoses are summarized in Table 47. All levels below limit quantifiable(blq<0.5 ng/mL) were treated as zero for purposes of pharmacokineticanalysis.

The concentrations of d-amphetamine and L-lysine-d-amphetamine intactconjugate following administration of L-lysine-d-amphetamine at the lowand high dose for each individual subject as well as pharmacokineticparameters are presented in Tables 48-51. The concentrations ofd-amphetamine following administration of Adderall XR® or DexadrineSpansule® for each individual subject as well as pharmacokineticparameters are presented in Tables 52 and 53, respectively.Concentration-time curves showing L-lysine-d-amphetamine intactconjugate and d-amphetamine (ng/mL, FIGS. 52A and 53A and uM, FIGS. 52Band 53B) are presented in FIGS. 52 and 53. Extended release ofd-amphetamine from L-lysine-d-amphetamine was observed for both dosesand pharmacokinetic parameters (C_(max) and AUC) were proportional todose when the lower and higher dose results were compared (Table 43, 50and 54; FIGS. 52 and 53). Significant levels of d-amphetamine were notobserved until one-hour post administration. Only small amounts (1.6 and2.0 percent of total drug absorption, respectively for 25 and 75 mgdoses; AUC_(inf)-molar basis) of L-lysine-d-amphetamine intact conjugatewere detected with levels peaking at about one hour Table 49 and 51).The small amount of intact conjugate absorbed was rapidly and completelyeliminated with no detectable concentrations present by five hours evenat the highest dose.

In a cross-over design (identical subjects received Adderall XR® dosesfollowing a 7-day washout period), the higher L-lysine-d-amphetaminedose was compared to an equivalent dose of Adderall XR®. Adderall XR® isa once-daily extended release treatment for ADHD that contains a mixtureof d-amphetamine and l-amphetamine salts (equal amounts of d-amphetaminesulfate, d-/l-amphetamine sulfate, d-amphetamine saccharate, andd-/l-amphetamine aspartate). An equivalent dose of extended releaseDexadrine Spansule® (contains extended release formulation ofd-amphetamine sulfate) was also included in the study. As observed inpharmacokinetic studies in rats, oral administration ofL-lysine-d-amphetamine resulted in d-amphetamine concentration-timecurves similar to those of Adderall XR® and Dexadrine Spansule® (FIGS.54 and 55). The bioavailability (AUC_(inf)) of d-amphetamine followingadministration of L-lysine-d-amphetamine was approximately equivalent toboth extended release amphetamine products (Table 54). Over the courseof twelve hours, typically the time needed for effective once-dailytreatment of ADHD, the bioavailability for L-lysine-d-amphetamine wasapproximately equivalent to that of Adderall XR® (d-amphetamine plusl-amphetamine levels) and over twenty percent higher than that ofDexadrine Spansule®. Based on the results of this clinical study,L-lysine-d-amphetamine would be an effective once-daily treatment forADHD. Moreover, L-lysine-d-amphetamine afforded similar pharmacokineticsin humans and animal models, namely, delayed release of d-amphetamineresulting in extended release kinetics. Based on these observationsL-lysine-d-amphetamine should also have abuse-resistant properties inhumans.

TABLE 48 Individual Subject d-amphetamine Concentrations andPharmacokinetic Parameters Following Oral Administration of a 25 mg Doseof L-lysine-d-amphetamine to Humans. Subject Subject Subject SubjectSubject Subject Subject Subject Subject Subject 102 103 105 107 110 112113 116 117 120 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 0 0 0  0.5 0 0 0.625 0 0 0 0 0.78 0.769 0 0.2 0.4 162.1 1 4.29 2.95 8.67 3.368.33 1.1 10 10.5 14 3.15 6.6 4.2 63.6   1.5 10 12.7 16 13.8 21.4 3.9424.7 19.5 24 15.1 16.1 6.5 40.3 2 16.3 18.4 17 21 25.9 9.29 30.9 23.6 3021.7 21.4 6.6 30.8 3 16.5 19.6 16.7 26.1 27 17.7 30.2 23.5 27.6 28.923.4 5.3 22.7 4 23.9 18.8 14.1 24.5 30.1 17.9 33.2 21.2 24.7 25.3 23.45.7 24.3 5 21.2 18.9 14.6 21.6 22.6 17.2 27 20 20.2 24.2 20.8 3.5 16.9 621.8 18 12.5 21.6 23.7 15.7 25.8 18.2 20.3 20.5 19.8 3.9 19.6 7 18.915.8 12.1 17.8 20.6 14.5 26.6 21 18.3 21.8 18.7 4.1 21.9 8 19.3 16.610.4 17.9 20 14.2 25.7 13.6 18.8 20.1 17.7 4.2 24.1 10  18.8 13.6 9.815.3 19.3 13.7 22.4 15.1 15.3 15.9 15.9 3.5 22.1 12  15.8 12.6 6.92 11.515.8 11.2 17.9 12 13.7 15.2 13.3 3.1 23.6 16  13.4 10.5 6.56 9.53 14.310.7 12.5 10.3 10 13 11.1 2.3 20.5 24  8.03 5.81 2.65 4.9 5.8 5.9 6.576.13 4.52 5.45 5.6 1.4 25.1 48  1.57 1.36 0 1.26 0.795 1.44 1.24 1.230.864 0.586 1.0 0.5 46.1 72  0 0 0 0 0 0 0 0 0 0 0 0 0 ParameterAUC_(0-12 h) 204.0 177.4 140.4 204.9 242.7 152.4 284.6 199.2 225.5 223.3205.4 42.5 20.7 (ng · h/mL) AUC_(last) (ng · h/mL) 463.3 375.1 201.4378.5 462.7 350.7 515.2 397.9 395.7 426.1 396.7 84.8 21.4 AUC_(inf) (ng· h/mL) 486.7 397.1 233.5 398.8 472 374 532.5 416.4 407 432.2 415.0 80.119.3 C_(max) (ng/mL) 23.9 19.6 17 26.1 30.1 17.9 33.2 23.6 30 28.9 25.05.6 22.3 T_(max) (hours) 4 3 2 3 4 4 4 2 2 3 3.1 0.876 28.2 T_(1/2)(hours) 10.32 11.18 8.36 11.18 8.16 11.22 9.68 10.43 9.06 7.22 9.68 1.4314.7

TABLE 49 Individual Subject L-lysine-d-amphetamine Intact ConjugateConcentrations and Pharmacokinetic Parameters Following OralAdministration of a 25 mg Dose of L-lysine-d-amphetamine to Humans.Subject Subject Subject Subject Subject Subject Subject Subject SubjectSubject 102 103 105 107 110 112 113 116 117 120 Mean SD CV % Time Hours0 0 0 0 0 0 0 0 0 0 0 0 0 0   0.5 4.1 5.5 10.0 0.0 3.6 0.0 9.2 9.6 8.90.0 5.1 4.2 82.0 1 9.2 11.2 15.2 12.5 9.1 2.7 20.1 10.5 10.8 10.9 11.24.5 39.7   1.5 4.0 4.4 6.1 7.5 3.6 6.2 6.6 2.8 4.2 8.4 5.4 1.8 34.1 22.1 1.4 2.5 2.9 1.9 4.0 2.3 0 1.7 3.1 2.2 1.1 48.8 3 0 0 0 0 0 0 0 0 0 00 0 0 4 0 0 0 0 0 0 0 0 0 0 0 0 0 5 0 0 0 0 0 0 0 0 0 0 0 0 0 6 0 0 0 00 0 0 0 0 0 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 0 0 8 0 0 0 0 0 0 0 0 0 0 0 00 10  0 0 0 0 0 0 0 0 0 0 0 0 0 12  0 0 0 0 0 0 0 0 0 0 0 0 0 16  0 0 00 0 0 0 0 0 0 0 0 0 24  0 0 0 0 0 0 0 0 0 0 0 0 0 48  0 0 0 0 0 0 0 0 00 0 0 0 72  0 0 0 0 0 0 0 0 0 0 0 0 0 Parameter AUC_(last) 9.18 10.9516.31 10.68 8.583 5.439 18.51 10.77 12.35 10.41 11.32 3.74 33.1 (ng ·h/mL) AUC_(inf) 10.62 11.64 17.66 12.65 9.759 — 19.56 — 13.3 12.83 13.503.40 25.2 (ng · h/mL) C_(max) (ng/mL) 9.18 11.2 15.2 12.5 9.05 6.18 20.110.5 10.8 10.9 11.56 3.80 32.9 T_(max) (hours) 1 1 1 1 1 1.5 1 1 1 11.05 0.16 15.1 T_(1/2) (hours) 0.47 0.34 0.38 0.47 0.44 — 0.32 — 0.380.55 0.419 0.077 18.5

TABLE 50 Individual Subject d-amphetamine Concentrations andPharmacokinetic Parameters Following Oral Administration of a 75 mg Doseof L-lysine-d-amphetamine to Humans. Subject Subject Subject SubjectSubject Subject Subject Subject Subject Subject 101 104 106 108 109 111114 115 118 119 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 0 0 0  0.5 0 0.748 0.506 0 0 0.779 0.525 0 3 1.85 0.7 1.0 132.2 1 11.9 14.412.6 7.26 5.9 10.3 7.2 23.1 23 27.9 14.4 7.7 53.6   1.5 40.3 34.6 30.422.8 19.3 38.4 19 52.8 51.5 55.8 36.5 13.8 37.8 2 84.6 48.9 68.2 34.832.7 57.2 33.1 91.3 61.7 70.4 58.3 21.0 36.0 3 72.9 64.3 55.7 60.3 62.361.1 44.8 95.8 62.1 83.6 66.3 14.5 21.9 4 84.6 65.3 58.8 51.1 77.9 63.347.6 89.2 54.2 86 67.8 15.5 22.8 5 65 55.6 60.2 74 83.9 59.1 56.9 77.754.9 82.8 67.0 11.5 17.2 6 71 53.5 49.4 51.5 78.3 50.8 55.1 68.8 52.9 6459.5 10.2 17.1 7 53.8 55.7 52.9 69.5 73.1 52.9 55.9 71.2 45.1 74.6 60.510.5 17.4 8 63.7 40.3 47.3 45.7 72.2 46.5 54.2 61.1 44.3 66.2 54.2 10.920.2 10  43.7 41.7 37 58.4 67 44.3 48.4 68 34.1 55.9 49.9 11.9 24.0 12 46.4 26.1 36.7 37.4 49.9 32.4 37.1 54.1 34.5 45.1 40.0 8.6 21.6 16  35.422.2 25.7 48 44.9 24.3 28.9 44.7 31.7 34.5 34.0 9.2 27.1 24  16.4 11.414.9 13.2 18.4 16.8 20.5 21.7 15.7 18.1 16.7 3.1 18.8 48  2.74 2.14 4.172.73 3.75 4.81 2.81 4.26 3.36 3.4 0.9 25.9 72  0 0 0 1.07 0.661 0.6871.49 0 0 0.553 0.4 0.5 120.2 Parameter AUC_(0-12 h) 666.2 525.9 531.6570.3 704.8 545.6 513.7 790.9 523.4 742.8 611.5 104.5 17.1 (ng · h/mL)AUC_(last) 1266 918.7 1031 1257 1442 1123 1223 1549 1143 1417 1237.0194.0 15.7 (ng · h/mL) AUC_(inf) 1301 948.3 1072 1278 1451 1133 12511582 1154 1425 1259.5 191.3 15.2 (ng · h/mL) C_(max) (ng/mL) 84.6 65.368.2 74 83.9 63.3 56.9 95.8 62.1 86 74.0 12.9 17.4 T_(max) (hours) 4 4 25 5 4 5 3 3 4 3.9 1.0 25.5 T_(1/2) (hours) 8.78 9.59 10.02 13.26 9.2410.41 12.8 8.05 10.92 9.47 10.3 1.7 16.3

TABLE 51 Individual Subject L-lysine-d-amphetamine Intact ConjugateConcentrations and Pharmacokinetic Parameters Following OralAdministration of a 75 mg Dose of L-lysine-d-amphetamine to Humans.Subject Subject Subject Subject Subject Subject Subject Subject SubjectSubject 101 104 106 108 109 111 114 115 118 119 Mean SD CV % Time Hours0 0 0 0 0 0 0 0 0 0 0 0 0 0   0.5 10.4 22.6 6.92 10.3 0 9.21 7.88 14.587.8 35.5 20.5 25.6 124.7 1 48 40.5 29 41.5 21.2 30.8 23.4 127 88.9 80.153.0 34.6 65.2   1.5 28.4 15.7 16.1 20.3 26.5 19 12.7 38.7 28.6 38 24.49.2 37.5 2 8.87 5.53 4.91 9 18.1 5.62 6.29 12.1 9.75 11.3 9.1 4.0 44.0 32.15 1.29 1.76 1.82 10.6 0 2.31 2.57 1.73 1.73 2.6 2.9 111.6 4 0 0 1.090 4.65 0 1.53 1.01 0 0 0.8 1.5 176.9 5 0 0 0 0 0 0 0 0 0 0 0 0 0 6 0 0 00 0 0 0 0 0 0 0 0 0 7 0 0 0 0 0 0 0 0 0 0 0 0 0 8 0 0 0 0 0 0 0 0 0 0 00 0 10  0 0 0 0 0 0 0 0 0 0 0 0 0 12  0 0 0 0 0 0 0 0 0 0 0 0 0 16  0 00 0 0 0 0 0 0 0 0 0 0 24  0 0 0 0 0 0 0 0 0 0 0 0 0 48  0 0 0 0 0 0 0 00 0 0 0 0 72  0 0 0 0 0 0 0 0 0 0 0 0 0 Parameter AUC_(last) 51.2 44.232.0 43.7 50.4 30.9 29.8 102.1 110.8 86.1 58.1 30.2 52.0 (ng · h/mL)AUC_(inf) 52.5 45.0 33.0 44.9 52.3 34.2 31.4 102.9 111.7 87.0 59.5 29.950.2 (ng · h/mL) C_(max) 48.0 40.5 29.0 41.5 26.5 30.8 23.4 127.0 88.980.1 53.6 34.1 63.6 (ng/mL) T_(max) (hours) 1 1 1 1 1.5 1 1 1 1 1 1.050.16 15.1 T_(1/2) (hours) 0.43 0.4 0.61 0.43 1.02 0.41 0.75 0.56 0.380.35 0.534 0.211 39.6

TABLE 52 Individual Subject d-amphetamine Concentrations andPharmacokinetic Parameters Following Oral Administration of a 35 mg Doseof Adderall XR ® (equivalent to 75 mg dose of L-lysine-d-amphetaminebased on amphetamine base content) to Humans. Subject Subject SubjectSubject Subject Subject Subject Subject Subject Subject 101 104 106 108109 111 114 115 118 119 Mean SD CV % Time Hours 0 0 0 0 0 0 0 0 0 0 0 00 0   0.5 7.9 2.3 2.8 0.6 2.2 5.7 0 16 2.3 5.3 4.5 4.7 104.3 1 37.6 28.923.3 13.7 29.8 38.2 17.9 46.2 28.8 48.8 31.3 11.5 36.6   1.5 49.9 42.331.1 23.7 39.1 34.4 30.8 65.4 34.1 53 40.4 12.5 31.0 2 65.9 45.8 29.237.4 46.2 65.4 40 64.4 37 67.8 49.9 14.6 29.2 3 95.3 51.7 36.7 23.6 64.762.9 44.7 56.5 31.1 64.8 53.2 20.7 38.9 4 83.7 73.3 56.7 40 67 76.6 56.353.1 33.5 73.3 61.4 16.3 26.6 5 77.4 75.2 71.6 62.1 75.9 76.4 51.5 61.456.8 82.4 69.1 10.3 14.9 6 71.5 72.1 64 59.8 66.9 63.5 56.8 59.8 58.785.7 65.9 8.7 13.2 7 72.3 63.6 71 57.9 70.6 69.7 51.9 48.1 53.7 79.763.9 10.5 16.4 8 60.4 57.1 53.8 53 72 66.9 56.2 56.4 51.7 66.7 59.4 6.911.6 10  50.4 45.5 53 50.7 67.6 57.4 49.1 66.6 48 71.3 56.0 9.3 16.6 12 42.5 41.3 45.4 32.9 53.1 46 37.3 74.7 42.2 60.2 47.6 12.2 25.7 16  31.129.6 35.7 39 45.2 33.9 34.3 64.9 29 40.5 38.3 10.6 27.7 24  14.9 15.122.1 19.5 21.7 21.2 20.7 35.7 17.9 20.5 20.9 5.8 27.7 48  2.5 4.2 3.85.9 5.4 3.8 7.3 5.1 3.9 3 4.5 1.4 32.1 72  0 0.3 1 1 0.3 1.1 2.7 0.3 0 00.7 0.8 124.7 Parameter AUC_(0-12 h) 731.2 625.0 582.6 504.3 711.6 698.5535.4 683.5 509.8 793.2 637.5 101.1 15.9 (ng · h/mL) AUC_(last) 12701230 1343 1269 1568 1436 1354 1920 1101 1520 1401.1 229.0 16.3 (ng ·h/mL) AUC_(inf) 1301 1234 1358 1286 1571 1454 1418 1923 1164 1557 1426.6218.9 15.3 (ng · h/mL) C_(max) 95.3 75.2 71.5 62 75.9 76.5 56.8 74.758.8 85.8 73.3 11.9 16.3 (ng/mL) T_(max) (hours) 3 5 5 5 5 4 6 12 6 65.70 2.41 42.2 T_(1/2) (hours) 8.65 9.01 10.57 11.58 8.37 10.78 16.47.25 11.05 8.54 10.22 2.59 25.3

TABLE 53 Individual Subject d-amphetamine Concentrations andPharmacokinetic Parameters Following Oral Administration of a 30 mg Doseof Dexadrine Spansule ® (equivalent to 75 mg dose ofL-lysine-d-amphetamine based on amphetamine base content) to Humans.Subject Subject Subject Subject Subject Subject Subject Subject SubjectSubject 102 103 105 107 110 112 113 116 117 120 Mean SD CV % Time Hours0 0 0 0 0 0 0 0 0 0 0 0 0 0   0.5 1.2 2.68 1.37 1.4 1.16 2.36 6.75 2.634.95 3.43 2.8 1.8 65.5 1 14.8 26.5 16.7 21.4 25.2 12.7 33.1 22.3 26 21.522.0 6.1 27.8   1.5 24.2 36.9 23.2 28.5 37.2 21.3 42.4 29.2 33.7 39.231.6 7.3 23.2 2 28.6 43.4 27.3 34.6 38.5 27.6 46.2 31.3 38.5 42 35.8 6.919.4 3 27.4 37.3 30.6 40.1 41.7 30.9 52 36.5 42.9 60.1 40.0 10.0 25.2 427.1 44.1 33.5 48.7 45.2 34.7 49.1 40.7 42.4 53.2 41.9 8.1 19.2 5 35.153 40.2 43.4 46.5 42.4 58.1 47 52.1 68.7 48.7 9.7 20.0 6 33.8 58.5 40.246.5 43.5 37.5 56.2 40 51 63 47.0 9.8 20.8 7 37.2 50.7 31.2 41.4 44.9 4257.8 43.6 51.6 65.7 46.6 10.1 21.7 8 35.9 54.3 34.9 45 45 36 58.7 41.853.9 59.2 46.5 9.5 20.4 10  33.1 49.1 34.3 35.5 45 37 51.4 38.9 46.360.1 43.1 8.8 20.4 12  34 51 28.6 34.1 40.8 32.6 51.6 37.7 38.1 50.939.9 8.4 21.1 16  30.2 40.8 25.2 28 33 25.8 41 26.8 29.6 44.9 32.5 7.122.0 24  20.5 27.8 18.2 19.5 17.1 17.8 22.5 19.1 15.5 27.3 20.5 4.2 20.348  3.83 6.89 3.7 5.11 2.56 4.31 6.51 4.43 2.77 5.47 4.6 1.4 31.8 72 0.715 1.63 1 1.7 0 0.622 1.29 1.22 0 1.31 0.9 0.6 64.0 ParameterAUC_(0-12 h) 356.2 539.8 366.4 444.3 480.8 387.0 591.4 436.5 512.8 634.2474.9 94.7 19.9 (ng · h/mL) AUC_(last) 1033 1517 966 1135 1065 1003 14731100 1048 1589 1193 236 19.8 (ng · h/mL) AUC_(inf) 1043 1544 983.5 11681097 1013 1495 1121 1085 1610 1216 238 19.5 (ng · h/mL) C_(max) (ng/mL)37.2 58.5 40.2 48.7 46.5 42.4 58.7 47 53.9 68.7 50.18 9.74 19.4 T_(max)(hours) 7 6 5 4 5 5 8 5 8 5 5.80 1.40 24.1 T_(1/2) (hours) 9.92 11.7412.07 13.8 8.7 10.76 11.47 12.23 9.36 10.92 11.10 1.50 13.6

TABLE 54 Pharmacokinetic Parameters of Amphetamine Following OralAdministration of L-lysine-d-amphetamine. Adderall XR ® or DexadrineSpansule ®. Drug L-lysine- L-lysine- d- d- amphetamine amphetamineAdderall Dexadrine Parameter 25 mg Percent¹ 75 mg Percent¹ XR ® Percent¹Spansule ® Percent¹ AUC_(0-12 h) 205.4 33.6 611.5 100 637.5 104 474.9 78(ng · h/mL) AUC_(last) (ng · h/mL) 396.7 31.5 1237 100 1401.1 113 119396 AUC_(inf) (ng · h/mL) 415.0 32.9 1260 100 1427 113 1216 97 C_(max)(ng/mL) 25.0 33.8 74 100 73.3 99 50.2 68 T_(max) (hours) 3.1 79.5 3.9100 5.7 146 5.8 149 T_(1/2) (hours) 9.68 94 10.3 100 10.22 99 11.1 108¹Percent relative to L-lysine-d-amphetamine 75 mg dose

It will be understood that the specific embodiments of the inventionshown and described herein are exemplary only. Numerous variations,changes, substitutions and equivalents will occur to those skilled inthe art without departing from the spirit and scope of the invention. Inparticular, the terms used in this application should be read broadly inlight of similar terms used in the related applications. Accordingly, itis intended that all subject matter described herein and shown in theaccompanying drawings be regarded as illustrative only and not in alimiting sense and that the scope of the invention be solely determinedby the appended claims.

1.-35. (canceled)
 36. A method of treating a subject with obesity, themethod comprising orally administering to the subject, a therapeuticallyeffective amount of L-lysine-d-amphetamine or a pharmaceuticallyacceptable salt thereof.
 37. The method of claim 36, wherein theL-lysine-d-amphetamine or a pharmaceutically acceptable salt thereof isin a dosage form selected from the group consisting of a tablet, acapsule, a caplet, an oral solution, or an oral suspension.
 38. Themethod of claim 36, wherein the L-lysine-d-amphetamine is in the form ofa salt.
 39. The method of claim 38, wherein the salt is a mesylate salt.40. The method of claim 38, wherein the salt is a hydrochloride salt.41. The method of claim 36, wherein the therapeutically effective amountis an amount sufficient to provide a therapeutically bioequivalent AUCwhen compared to amphetamine alone, but which does not provide a C_(max)which results in euphoria.
 42. The method of claim 37, wherein thetherapeutically effective amount in the dosage form is from about 5 mgto about 500 mg of the L-lysine-d-amphetamine or a salt thereof.
 43. Themethod as defined in claim 42, wherein the dosage form is administeredone or more times per 24-hour period.